Upon physical interaction, cyclin-dependent kinase 5 (Cdk5) and p25 forms hyperactivated Cdk5/p25 complex and causes severe neuropathological aberrations including hyperphosphorylated tau-mediated neurofibrillary tangles formation, Alzheimer’s symptoms, and neuronal death. Therefore, the inhibition of Cdk5/p25 complex may relieve p-tau-mediated Alzheimer’s pathology. Herein, computational simulations have identified pyrrolidine-2,3-dione derivatives as novel inhibitors of Cdk5/p25 complex. A ligand-based pharmacophore was designed and employed as 3D query to retrieve drug-like molecules from chemical databases. By molecular docking, drug-like molecules obtaining dock score > 67.67 (Goldcore of the reference compound) were identified. Molecular dynamics simulation and binding free energy calculation retrieved four pyrrolidine-2,3-dione derivatives as novel candidate inhibitors of Cdk5/p25. The root means square deviation of Cdk5/p25 in complex with candidate inhibitors obtained an average value of ~2.15 Å during the 30 ns simulation period. Molecular interactions analysis suggested that each inhibitor occupied the ATP-binding site of Cdk5/p25 and formed stable interactions. Finally, the binding free energy estimation suggested that each inhibitor had lowest binding energy than the reference compound (−113.10 kJ/mol) to recapitulate their strong binding with Cdk5/p25. Overall, these inhibitors could mitigate tau-mediated Alzheimer’s phenotype.Mechanistically, neurotoxic insults provoke Ca Cyclin-dependent kinases (Cdks) play essential roles in post-translational modification of several proteins, thereby modulating their physiological fate. Cdks belong to the CMGC  sub-family protein kinases and expedite the transfer of γ-phosphate from adenosine triphosphate (ATP) to peptide substrate(s) . Cyclin-−/− mice die just before or after birth due to abnormal corticogenesis, neuronal pathology, and detained cerebral cortex development [Cdk5 regulates various neuronal functions including physiological development of the central nervous system (CNS), neurogenesis, neuronal migration, synaptic remodeling, synaptic activity, and learning and memory ,8,9. Mutelopment . Cdk5 alelopment . Additioelopment .To date, numerous studies have supported the role of Cdk5 in several pathological phenotypes including cancer, neurodegenerative diseases, and ischemia. Abnormal regulation of Cdk5 by truncated activators (p25 and p29) contribute to neurodegeneration and has been implicated in Alzheimer’s diseases (AD), amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Niemann–Pick type C disease, and Ischemia ,14. CleaTo this end, the pharmacological inhibition and/or targeted knockdown of Cdk5 relieved neurotoxicity and tau pathology . LikewisNonetheless, available Cdk5 inhibitors may not be clinically translatable drugs to potentiate treatment platform in human neurological disorders . ConsequMinimize Ligands module, implanted in DS. Thereafter, Common Feature Pharmacophore Generation module of DS was employed to generate pharmacophore hypotheses. Common Feature Pharmacophore Generation module uses the HipHop algorithm and determines the atom-based critical common chemical features of all the compounds of a given training set to create 3D-pharmacophore models [Best and Flexible mode, and minimum and maximum number of features were set to 3 and 6, respectively [The inhibitors of Cdk5, investigated by the same biological assays, were identified from literature survey ,33,34. Te models . The proectively . FinallyLigand Pharmacophore Mapping protocol, implanted in DS. The pharmacophore-mapped molecules were used as input values and parameters like goodness score (GH), enrichment factor (EF), percent ratio of actives (%A), percent yield of actives (%Y), false negative, and false positive were measured by equations:a: number of actives in hit list; Ht: total number of hits retrieved by pharmacophore; D: total number of compounds in decoy test set; A: total number of actives in decoy test set.Pharmacophore validation evaluates the potentiality of pharmacophore to identify target compounds . PharmacLipinski’s Rule of Five and ADMET Descriptors modules of DS to identify the drug-like compounds. Lipinski’s rule of five evaluates the physiochemical properties of chemical compounds [Since, drug-like properties are pre-requisites for a chemical compound to be used as a drug; therefore, the selected chemical databases  were filtered by ompounds . The ADMLigand Pharmacophore Mapping protocol, implanted in DS. The screening was carried out under the Best/Flexible parameterization environment at the Maximum Omitted Features “0”, which ensures the mapping of hit molecules onto all the features of the pharmacophore [The validated pharmacophore was employed as a 3D query and each drug-like database was screened by acophore .Genetic Optimization of Ligand Docking  package allows full flexibility of ligands and partial flexibility of protein; therefore, it estimates more reliable calculations in docking simulations [add hydrogen) was used to add hydrogen atoms to Cdk5/p25. The ligand binding site of Cdk5 was defined within a radius of 7.00 Å of the inbound inhibitor (Roscovitine). The Goldscore and ASP  scoring functions were used as the default scoring and rescoring functions, respectively [Molecular docking is a state-of-the-art approach in computational biology to identify and evaluate molecular interactions between the ligands and receptors ,43,44,45ulations . Herein,ectively . The Golectively . The CdkGroningen Machine for Chemical Simulation (GROMACS v5.1.4) package [LINCS algorithm which allowed 2 fs time step in all simulations. During simulation, V-rescale thermostat and Parrinello–Rahman barostat were employed to sustain the temperature and pressure at 300 K and 1.0 bar, respectively.From docking analysis, the true positive candidate inhibitors (successfully docked candidate hits) of Cdk5/p25 were subjected to molecular dynamics simulation to explore their stability and mechanism of interaction under the simulated physiological environment. For each drug-like molecule, independent simulation system was prepared in  package . The par package , whereas package . Each sy package . Periodibind) of the reference compound and final candidate hits with Cdk5/p25 were calculated by molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) method [bind) in solution is defined as:The binding free energies (ΔG) method . The binA molecular dynamics simulation generates an ensemble of time-equidistance conformations. The free energy term is calculated as an average over the representative structures as:MM is defined as:intE indicates bond, angle, and torsional angle energies, and coulE and LJE display the intermolecular electrostatic and van der Waals energies, respectively.The energetic term Esolv is the combination of Gpolar and nonpolar contribution, Gnonpolar:The solvation term Gnonpolar is proportional to the solvent accessible surface area (SASA):−1Å−2 and β = 3.849 kJ mol−1.The non-polar contribution GHerein, an ensemble of thirty time-equidistant snapshots of candidate inhibitors in complex with Cdk5/p25 were taken from the entire MD trajectories and the binding free energy calculations were performed.Ligand-based pharmacophore modeling is a well-established approach in computational drug discovery processes. Herein, eight chemically diverse inhibitors of Cdk5 were collected from literature mining ,33,34. TFeature Mapping module, implanted in DS. It was observed that the training set compounds possess hydrogen bond acceptor (HBA), hydrogen bond donor (HBD), hydrophobic (HYP), hydrophobic aromatic (HYA) and ring aromatic (RAR). Afterwards, the Common Feature Pharmacophore Generation module of DS was employed and ten pharmacophore models were generated. Our results showed that the ranking scores of hypotheses ranged from 55.82 to 75.17 kcal/mol  features, one hydrogen bond donor (HBD) feature, and one hydrophobic (HYP) feature . Such coThe detailed study of Hypo1 suggested that polar interactions are the pre-dominant features and delineates the chemical space of the selected pharmacophore. The four features of Hypo1 was composed of two hydrogen bond acceptors (HBA), one hydrogen bond donor (HBD), and one hydrophobic (HYP) feature, respectively A. The spThe potentiality of Hypo1 was evaluated to differentiate between the active and inactive inhibitors of Cdk5/p25. A decoy test set was prepared, which was comprised of 40 active inhibitors of Cdk5 (A) and 668 inactive molecules of Cdk5 .t) were mapped, where 38 compounds were the active inhibitors (Ha) of Cdk5. Our results suggested that Hypo1 mapped 95% active inhibitors and obtained the highest percent yield of 82.6%  was used to profile the physiochemical properties of three chemical databases including NCI, Asinex, and Specs databases. According to ROF, a drug-like compound should be membrane permeable and could easily be absorbed if it has total number of hydrogen bond acceptors and hydrogen bond donors less than 10 and 5, and molecular weight and AlogP values are less than 500 Da and 5, respectively [A chemical compound is used as a drug-like molecule if its physiochemical, pharmacodynamics, and pharmacokinetics properties exhibit drug-like properties . Herein,ectively . Our resADMET Descriptors module in DS. Herein, the drug-like compounds were evaluated for their potentiality to cross blood-brain barrier, solubility, hepato-toxicity, cytochrome toxicity, and absorption  atoms of Cdk5/p25 suggested that all the systems were converged after 4 ns simulation  complex are given in Furthermore, the backbone RMSD of each system suggested that each system behaved normal during the entire simulation period B. SimilaSince, it has already been reported that small molecule inhibitors occupied the ATP-binding site of Cdk5/p25 ,59. TherFurthermore, we focused on polar interactions (H-bond formation) of hit(s) with the Cys83 residue of Cdk5, which has already been investigated as the potential mechanism of Cdk5/p25 inhibition ,59,62. ILikewise, we addressed whether H-bonds between the Cys83 of Cdk5 and the REF compound and/or hit molecules are consistent throughout the production phase. Fortunately, our results demonstrated that all the hit molecules showed stable consistency of the H-bond formation with the Cys83 residue of Cdk5 throughout the production phase B. Since,Since, non-polar interactions play a significant role in protein–drug interaction ,59,60,68Since, the REF compound and final hit molecules showed stable RMSD values and strong molecular interactions with Cdk5/p25. Therefore, we argued to identify the candidate hits with strong binding affinity towards Cdk5/p25. To this end, the binding free energy analysis of the REF compound and each hit molecule was carried out against the Cdk5/p25 complex. MM/PBSA is a well-established approach to explore binding free energy between the protein and ligand(s) . Our resBased on binding free energy analysis, our results proclaimed that the hit molecules had higher affinity towards Cdk5/p25 and may strongly inhibit Cdk5/p25. The ΔG values for all the tested compounds were in the range of −113.10 to −137.90 kJ/mol. The average binding free energy value of each candidate hit is given in α atoms and backbone atoms of Cdk5/p25, and Cdk5/p25 in complex with hit molecules revealed that all the candidate molecules obtained lowest RMSD value (<2.5 Å) and behaved stable during the production phase of simulation. Furthermore, the final hit molecules occupied the ATP-binding site of Cdk5/p25 and formed consistent hydrogen bonds with the Cys83 residue the ATP-binding site of Cdk5/p25. Finally, MM/PBSA approach suggested that each candidate inhibitor had lowest binding free energy with the Cdk5/p25 complex to recapitulate their strong binding. Overall, we suggest that the inhibition of Cdk5/p25 by the newly identified pyrrolidine-2,3-dione derivatives may alleviate the tau-associated Alzheimer’s pathology.The abnormal production of p25-mediated hyperactive cyclin-dependent kinase 5 (Cdk5) results in aberrant hyperphophorylation of tau protein, neuroinflammation, and Alzheimer’s phenotypes. Therefore, the inhibition of Cdk5/p25 complex may relieve the aberrant phosphorylation of tau protein to mitigate AD pathology. Herein, computational simulations have identified pyrrolidine-2,3-dione derivatives as the candidate inhibitors of Cdk5/p25 complex to alleviate AD pathologies. A ligand-based pharmacophore was generated and validated by decoy test set method. The validated pharmacophore retrieved drug-like compounds from chemical databases like NCI, Asinex, and Specs databases. Molecular docking approach filtered the drug-like molecules and identified the candidate hit compounds of Cdk5/p25. Molecular dynamics simulation and binding free energy calculation retrieved four hit molecules as the best candidate inhibitors of Cdk5/p25 complex. The root mean square deviation (RMSD) analysis of C


UBE3A gene is imprinted in the brain with preferential maternal-specific expression particularly in the neuron and loss of activity of the maternally inherited UBE3A causes Angelman syndrome (AS), characterized by severe mental retardation, lack of speech, seizures and autistic features. Interestingly, duplication, triplication, or gain-of-function mutations in the UBE3A gene are also linked with autism clinically distinguished by social impairments and stereotyped behaviors. These findings indicate that the expression and activity of UBE3A must be tightly regulated during brain development and UBE3A might be playing a crucial role in controlling synaptic function and plasticity through proteasome-mediated degradation as well as transcriptional regulation of its target proteins. In fact, several recent reports demonstrated the role of UBE3A in the modulation of synaptic function and plasticity. This review focuses on the critical role of UBE3A in regulating the synaptic function and how its altered activity is associated with autism.UBE3A is a dual function protein consisting of ubiquitin ligase as well as transcriptional co-activator function.  With the extensive study in multiple new cases, the Diagnostic and Statistical Manual of Mental Disorders (DSM-V) updated this definition in 2013, wherein, delayed language development was removed from the list of criteria and autism covered a host of diseases redefining itself as an autism spectrum disorder (ASD). First reports of autism were made by Leo Kanner and Hans Asperger in 1943–44. Both Kanner and Asperger defined a set of children withdrawn in their own world and failing to express themselves in social settings. Kanner termed this condition  Kanner, .in utero exposure to alcohol, recreational drugs, pesticides, prenatal valproate exposure, nutritional deficiency, and maternal infections may hamper neural development leading to ASD   that are known to be associated with Timothy syndrome  , intracellular trans-membrane proteins of the Golgi complex (POMGNT1), and genes responsible for proper cerebellar function (SYNE1) are important few names among multitude of genetic factors that may cause ASD , a neurodevelopmental disorder with severe mental retardation along with autistic features  receptor subunits β3, α5, γ3, and CYFIPI (cytoplasmic FMRP-interacting protein 1) are also reported in autism  domain family. As a part of the ubiquitin proteasome system, ubiquitin ligases (such as UBE3A) selectively target specific proteins for their ubiquitination and subsequent degradation by proteasome  generated by Jiang et al reproduced many characteristic features of AS including cognitive and motor deficits, audiogenic seizure, anxiety-like behavior, disturbances in circadian clock, and sleep homeostasis  recombinant technique. They inserted the 162 kb fragment of mouse chromosome 7 which contained the entire 78 kb coding sequence of Ube3a gene as well as its 63 kb 5′ and 21 kb 3′ extragenic sequence into FVB embryos, which were subsequently bred to produce single or double copy transgenic mice  had impaired social behavior and communication along with increase in repetitive behavior, which is the hallmark of autism. And similar to the dup15 phenotype, the mouse with two fold increase in brain Ube3a (1xTg) had a weaker autism penetrance with few of behavioral deficits. This model suggested that Ube3a is a dose sensitive gene and it is required in triplication to reconstitute the full set of symptoms of autism in a mouse model system.The first mouse model of autism was developed by Nakatani et al. , where tNLGN3) showed increased GABAergic transmission, while mouse model for Rett syndrome (where neurons were depleted of MeCP2) showed decreased GABAergic transmission without affecting glutamatergic transmission  mice which were enriched for glutamatergic synaptic transmission. They found Cbln1, a gene which binds to Nrxn1/2/3 and postsynaptic GRID1/2 and is dose dependently affected by Ube3a. Both NRXN1/2/3 and GRID1/2 genes were found mutated in ASD in a network protein-protein interaction study  and Ube3a (2xTg) mouse models. Krishnan et al. also observed social deficits similar to Ube3a (2xTg) mouse in animals with Cbln1 deleted from VGLUT neurons specifically in the ventral tegmental area (VTA). They also reported that recurrent seizures reduced Cbln1 expression along with impaired sociability. This result was supported by an earlier report that showed down-regulation of Cbln1 mRNA by increased neuronal activity after status epilepticus  in excitatory neurons. Ube3a isoform 2 plays a key role in neural development hence was the isoform of choice for this model. Behavioral abnormalities in this mouse model were robust in contrast to the Ube3a (2xTg) mouse developed by Smith et al. Although it showed normal growth in the first 2 weeks post birth and had normal motor and exploratory abilities, this model had fewer USV  at both early and later stages of development. Overexpression of Ube3a isoform 2 also led to anxiety and cognitive impairments. This mouse model exhibited a unique feature of reduced seizure threshold as well as volume reduction and dysmorphia of the hippocampus which was in accordance with the phenotype of Dup15 patients, where 60% patients have seizures and are known to have heteropias and dysplasias  at the age of 4 months. She suffered global developmental delay, although she could later communicate with little vocabulary. She is social with friends at school and also can make good eye contact. Family history of the patient revealed learning disability, anxiety, and depression on the patient's maternal side. These reports strongly indicate that UBE3A is one of the major contributors to the autistic phenotype in cases with maternal interstitial 15q11–13 duplication.Lossifov et al. conducted a whole genome sequencing study, where they found autism proband with a T485A missense mutation in UBE3A. Yi et al. reported that chronic treatment with drugs targeting PKA, like forskolin and rolipram, can turn down UBE3A activity in neurons. This raises the possibility of targeting upstream regulators of UBE3A in order to rescue autistic symptoms   patients restored normal UBE3A mRNA levels using antitumor antibiotic methamycin (Germain et al., All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Prosopis species plays a prominent role as a food resource in arid areas. The aim of this work was the characterization of the phenolic antioxidants occurring in the pod mesocarp flour of Chilean Prosopis. Samples were collected in the Copiapo, Huasco and Elqui valleys from the north of Chile. The samples of P. chilensis flour exhibited a total phenolic content ranging between 0.82–2.57 g gallic acid equivalents/100 g fresh flour weight. The highest antioxidant activity, measured by the DPPH assay, was observed for samples from the Huasco valley. HPLC-MS/MS analysis allowed the tentative identification of eight anthocyanins and 13 phenolic compounds including flavonol glycosides, C-glycosyl flavones and ellagic acid derivatives. The antioxidant activity and the phenolic composition in the flour suggest that this ancient South American resource may have potential as a functional food.In South America, the mesocarp flour of  Prosopis, locally known as “algarrobo” in South America, are relevant food sources [Prosopis pods constitute a food source for humans and animals [Prosopis, including flour, sweets, syrup or fermented alcoholic beverages [Worldwide climate change and the long-lasting drought in northern Chile suggest that attention should be paid to native species that are adapted to arid environments. In arid and semi-arid lands, pods of the trees belonging to genus  sources . They we sources , Amerind sources ,4, Argen sources , and Chi sources ,7. Proso animals . Differeeverages ,8. Aftereverages ,4. The peverages  or is feeverages .Prosopis pods mesocarp flour was investigated for its food applications [Prosopis flour potential as food was revised by Felker et al. [et al. [In northern Chile and Peru, “algarrobo” pods were an important food source in pre-Hispanic times and they can be found in archeological sites and in burials . In Chilications  and has ications . The Pror et al.  and by C [et al. .Prosopis species were investigated for their alkaloid content and composition, as well as for some biological activities, including enzyme inhibition, antioxidant effect and DNA binding [Prosopis pods were also analysed in a study on the proximate composition and bioactivity of food plants consumed by Chilean Amerindians [et al. [Prosopis alba and P. nigra contains antioxidant and anti-inflammatory agents [Prosopis pods mesocarp flour.Chilean and Argentinean  binding ,16,17,18 binding ,20,21. Prindians ,9 as welrindians  was inve [et al. . Phenoliy agents , are noty agents  while bey agents . Lentil y agents . Methanoy agents . In contProsopis flours presented different hues according to the colour of the pods. Pods from Elqui valley showed sticky granules producing tacky flours when ground. The percent of mesocarp flour yield ranged from 36.7% flour to pod ratio for the deep purple Puquio sample, 31.8% for the Elqui valley and 3.3%–17.8% for the beige pods of P. chilensis collected in the Huasco valley, respectively. The sample from El Transito, Huasco Valley, presented a very low flour to pod ratio (3.3%). With thin pods, this sample is considered inedible by the local population [Six “algarrobo” pods samples were collected from the longitudinal valleys of Huasco, Elqui and Copiapó in northern Chile . The morpulation ,8. P. chilensis ranged from 0.10% to 2.08% for the Huasco and Elqui Valley samples (The range of phenolic compounds (TP) in flours was 0.82–2.57 g GAE per 100 g FFW. The higher values were from Alto del Carmen (2.75 g GAE/100 g FFW), Puquio (2.54 g GAE/100 g FFW) and Plaza de Pinte (2.11 g GAE/100 g FFW). Total flavonoid (TF) content in flour was low (0.17–0.56 g QE/100 g flour weight), and no correlation was observed between the TP and TF content. As TP and TF of the flour was low, samples were enriched in phenolics for antioxidant activity studies and phenolic profiling. The flour samples were extracted with MeOH and phenolics were retained on Amberlite XAD-7 to obtain the phenolic-enriched flour extract (PEFE). The highest PEFE was from the Copiapo Valley sample (4.26%). Lower PEFE values for the different samples of  samples .50 12.07 and 23.74 µg/mL, respectively). The same samples presented the highest activity in the FRAP assay, with values of 3.45 and 1.21 mM TE/g PEFE, respectively. In addition, the highest TEAC value was observed for the Alto del Carmen sample with a 3206.61 µM TE/g PEFE. There was a statistical correlation between the TP content and FRAP . Large variation in mesocarp flour yield and phenolic content was observed for P. chilensis. The close related species P. alba and P. nigra are common in the Chaco zone of South America and occur in the eastern Andean ranges of Argentina.The best antioxidant activity of the PEFE, measured by the DPPH discoloration assay, was found in the Huasco Valley samples: Alto del Carmen and Plaza de Pinte , with flavonoids accounting for 0.01 and 0.06 g/100 g DW, respectively. When the flour was extracted with water, the TP values increased to 0.40 and 0.41 g/100 g DW and TF to 0.03 and 0.13 g/100 g DW for P. alba and P. nigra, respectively. The TP (0.82–2.57 g GAE/100 g flour) and TF content (0.17–0.56 g QE/100 flour) of the Chilean Prosopis flour samples was higher than that of the Argentinian species. In a study by Cardozo et al. , TP in eProsopis species, Felker et al. [P. alba and P. nigra flour, respectively. According to the same author, TP for wheat bran ranges between 0.126–0.316 g GAE/100g DW or 0.27–0.35 g GAE/100 g DW while white wheat flour contains 0.0044–0.014 g GAE/100g DW [In a review on pod mesocarp flour of r et al.  refer ton. Anthocyanins were detected in the positive ion mode, while other phenolic compounds were analyzed in the negative ion mode. The representative HPLC-DAD chromatogram of PEFE from the Puquio sample is presented in 2 spectra of compounds 1–21 are presented in The composition of PEFE was assessed by HPLC-DAD-MS/MS1–3, 5–7, as detected in the DAD, show maxima in the 516–520 nm range, in agreement with anthocyanidins bearing a substituent at the 3-position [et al. [1–8, leading to a m/z ion at 287 amu for compounds 1, 2 and 5, m/z 301 for 3, 6 and 7, m/z 317 for 4 and 331 for 8. This is consistent with the occurrence of cyanidin, peonidin, petunidin and malvidin hexosides in the sample. The main compound was cyanidin 3-hexoside. A malonoyl unit was detected in compounds 5 and 6, assigned as cyanidin malonyl hexoside and peonidin malonyl hexoside, respectively. The visible spectra of compounds position ,29. Acco [et al.  3-glycosProsopis chilensis and are responsible, at least in part, of the dark purple colour of the pods. Compounds 9 and 11 presented the same pseudo molecular ion at 463 amu and loss of a hexose, leading to the ion at m/z 301. The elution time and UV spectrum of 11 is in agreement with a phenolic compound, consistent with ellagic acid. Both compounds differ in the identity of the hexose and were tentatively identified as ellagic acid hexosides. The compound 10 showed in the MS/MS spectrum the loss of a hexoside, leading to the aglycone at m/z 209, in agreement with hydroxyferulic acid hexoside [These anthocyanins have not been previously reported from hexoside .15–18, 20 and 21 show maxima in the range 349–354 nm, in agreement with the Band I of flavonol substituted at the 3-O-position. MS/MS analysis showed the loss of two 162 amu fragments (hexose) for compounds 15 and one hexose and a pentose for 16. The UV spectra of compounds 17, 18, 20 and 21, leading to aglycones with m/z 301 or 315, in agreement with quercetin  and quercetin methyl ether (17 and 21). The compounds were identified as quercetin glycosides 15, 16, 18 and 20 and quercetin methyl ether glycosides 17 and 21. Flavone C-glycosides show characteristic UV maxima around 330 nm (Band 1) and losses of 120 amu in the MS/MS spectra [12–14 and 19 presented UV and mass spectra in agreement with this group of compounds and were assigned as vicenin II isomers (12 and 13), schaftoside or isoschaftoside (14) and isovitexin (19). The C-glycosyl flavones schaftoside, isoschaftoside and vicenin II were recently identified as constituents of Prosopis pods syrup [Consecutive losses of rhamnose and hexose or rutinoside were observed for  spectra . Compounds syrup .Table 2Prosopis pod mesocarp flour extracts by HPLC-DAD-ESI-MS data. Anthocyanins (A) were detected in Puquio sample. Phenolics different than anthocyanins (B) were found in all phenolic-enriched pod mesocarp flour extracts.Tentative identification of anthocyanins and phenolics in phenolic-enriched O-glycosides, four flavone C-glycosides and a hydroxyferulic acid hexoside. Anthocyanins were detected only in the sample from the Copiapo Valley (Puquio). The compounds were cyanidin, peonidin, petunidin and malvidin derivatives, suggesting similarity with P. nigra [O-sambubioside, the 3-O-glucosides of delphinidin, cyanidin, pelargonidin and peonidin were described from the immature purple pods of yard-long beans [Prosopis pods until a recent study on P. alba and P. nigra mesocarp flour [Prosopis sample collected in Puquio. This place was a natural corridor that allowed crossing the Andes from the Copiapo Valley to the Argentinian provinces of Catamarca, La Rioja and Tucuman through the Paso San Francisco. As this was also a pre-Columbian route to cross the Andes and there is no previous records of P. nigra in Chile, we cannot rule out three possibilities: (i) that the distribution range of the species included part of the western Andean valleys; (ii) that the species was introduced in Chile by migrants or, (iii) that the sample is a high anthocyanin variety of P. chilensis. The pods as well as the starch from P. chilensis and P. flexuosa are frequently found in archeological remains in the dry valleys of Catamarca, Argentina [HPLC-DAD-MS/MS analysis of the PEFE allowed the tentative identification of eight anthocyanins, two ellagic acid glycosides, six flavonol P. nigra . AnthocyP. nigra , black rP. nigra , and purP. nigra . The antP. nigra  and cyanng beans . Despiterp flour . In the rgentina . C-glycosylflavones 12, 13 and 14 as the main compounds. In the Pinte, Plaza de Pinte and Alto del Carmen flours . 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) diammonium salt, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), potassium persulfate, sodium carbonate, FeCl3·6H2O, HPLC-grade methanol, acetonitrile and formic acid were purchased from Merck . Ultrapure water was obtained using a BarnstedEasyPure water filter .Folin-Ciocalteau phenol reagent, 2,4,6-tri(2-pyridyl)1,3,5-triazine (TPTZ) sodium acetate, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), quercetin, gallic acid and AlClProsopis chilensis by Dr. Patricio Peñailillo and voucher specimen were deposited at the Herbario de la Universidad de Talca. The collection places were as follows: Copiapo Valley: road to Paso San Francisco, near Puquio ; Huasco Valley: Alto del Carmen , El Transito ; road to Pinte  and Plaza de Pinte ; Elqui Valley: 30°06′39′′S, 70°29′58′′W. Samples were transported to the lab and kept at room temperature, according to the traditional storage indications. The air-dried pods were processed in a grinder to separate the seeds from the mesocarp flour. The traditional flour preparation was followed, using a mortar and pestle. The flour was sieved and weighed to establish the pod/mesocarp flour ratio. Pods flour was extracted with MeOH under sonication (2 × 3 min each time), in 1:10 flour to MeOH w/v ratio. The MeOH solution was filtered and taken to dryness under reduced pressure to afford the crude MeOH extract. The extracts were dissolved in water, filtered and adsorbed into Amberlite XAD-7, pre-treated as described in Jiménez-Aspee et al. [2O 7:3 (v/v) and the combined extracts of each sample were taken to dryness and lyophilized. The phenolic-enriched flour extracts (PEFE) were concentrated under reduced pressure and lyophilized for its analysis.Algarrobo pods were collected in the Copiapo, Huasco and Elqui valleys in February 2013 . The samet al. [2O (1:1). For TP, the Folin-Ciocalteu method was followed. The results are expressed as g gallic acid equivalents (GAE)/100 g fresh flour weight (FFW). For TF, the AlCl3 methodology was used. TF was expressed as g quercetin equivalents (QE)/100 g FFW. Absorbance of each solution were measured by spectrophotometer  at 725 and 510 nm, respectively, after 15 min of incubation at room temperature.The total phenolic (TP) and total flavonoid (TF) content was determined in the flour MeOH extract as described by Jiménez-Aspee et al. , with slv/v aqueous methanol at a final concentration of 300 μg/mL. Stock solutions were filtered and kept in the dark, and all analyses were performed on the same day.The antioxidant activity of the samples was determined by three assays, as described ,46. PEFE50 values (μg/mL), corresponding to the amount of extract that scavenges the radical concentration by 50%, were calculated using the OriginPro 8.0 software . DPPH discoloration assay was carried out with final concentrations of 100, 33 and 11 μg/mL. The DPPH solution was freshly prepared in methanol (20 mg/L) and mixed with the extract at the above given concentrations. Absorbance was measured at 517 nm in a universal microplate reader . SCFor the ferric reducing antioxidant power (FRAP) assay, a 300 μg/mL extract aliquot was mixed with warm FRAP solution and left to stand in the dark for 30 min. Absorbance was read at 593 nm using a spectrophotometer (Thermo Spectronic Helios Alfa) Results are expressed as mMoles Trolox equivalents (TE)/g extract.•+ with fresh standard (1 mM Trolox) or extract . Absorbances were read at 734 nm after 6 min of room temperature incubation using a spectrophotometer (Thermo Spectronic Helios Alfa). Results are expressed as μM Trolox equivalents/g extract.The Trolox-equivalent antioxidant capacity (TEAC) determinations were carried out by mixing ABTSet al. [The extracts were analysed by HPLC coupled to a diode array detector (HPLC-DAD) to set the conditions for HPLC-ESI-MS/MS studies. The HPLC system used for DAD analysis was a Shimadzu equipment  consisting of a LC-20AT pump, a SPD-M20A UV diode array detector, CTO-20AC column oven and a LabSolution software. A MultoHigh 100 RP 18–5µm (250 × 4.6 mm) column  maintained at 25 °C was used. Approximately 5 mg/mL of PEFE was filtered through a 0.45 µm filter  and injected into HPLC-DAD and HPLC-ESI-MS/MS. The compounds were monitored at 254, 330 and 535 nm, and UV spectra from 200 to 600 nm were recorded for peak characterization. The HPLC analysis was performed using a linear gradient solvent system as described by Quispe et al. . The flom/z 20–2200) and normal resolution . The trap parameters were set in ion charge control (ICC) using manufacturer default parameters, and maximum accumulation time of 200 ms. Collision induced dissociation (CID) was performed by collisions with helium background gas present in the trap and automatically controlled through Smart Frag option.The mass spectrometer consisted of a HPLC HP1100  connected through a split to the mass spectrometer Esquire 4000 Ion Trap LC/MS(n) system . Ionization was performed at 3000 V assisted by nitrogen as nebulizing gas at 24 psi and as drying gas at 365 °C and a flow rate of 6 L/min. Negative ions were detected using full scan  coupled to a MicroQTOF Q II . The HPLC system consisted in a micro vacuum degasser, binary pumps, an autosampler (40 μL sample loop), a thermostated column compartment and a diode array detector. The mass spectrometer equipped with an electrospray ion source and QTOF analyser, was used in MS and MS/MS mode for the structural analysis of phenolics. HPLC analyses were performed on a thermostated (40 °C) MultoHigh 100 RP 18–5µm (250 × 4.6 mm) column (5 μm) with a flow rate of 1.0 mL/min using a split to the detector. The solvents and ramp were the same as described for the ion trap equipment. 2 option. DAD analyses were carried out in the range between 200 and 700 nm. The identification of phenolic compounds in “algarrobo” pods meal was carried out by comparison of the spectral properties (UV and ESI-MS and MS/MS) of the compounds with literature data. ESI-MS detection was performed in negative and positive ion mode with mass acquisition between 100 and 1500 Da. Nitrogen was used as drying and nebulizer gas , and 180 °C for drying temperature. For MS/MS experiments, fragmentation was achieved by using Auto MSDeterminations of TP, TF, DPPH and FRAP, were performed in triplicate and results are expressed as mean values ± SD. For the TEAC assay, a curve was plotted for each sample and a correlation coefficient with 95% confidence limit was established. To assess the relationship between the antioxidant activities and the TP and TF content, Pearson’s correlation coefficients were calculated with 95% confidence. Statistical analysis was carried out using the software SPSS 14.0 for Windows.Prosopis pods. The phenolic composition and antioxidant properties of the Chilean Prosopis mesocarp flour supports its use as a functional food. Additional studies are required to compare the potential of the different flour sources in artisanal and commercial food products. A higher number of samples should be analyzed to have a better picture on the phenolic composition of Chilean Prosopis mesocarp flour. The main compounds in the PEFE were flavonoids. One sample contained cyanidin hexoside and other anthocyanins, being this the first report on the occurrence of anthocyanins in Chilean


Akkermansia muciniphila is important for the human health as the occurrence of the organism is inversely correlated with different metabolic disorders. The metabolism of the organism includes the degradation of intestinal mucins. Thus, the gut health‐promoting properties are not immediately obvious and mechanisms of bacteria‐host interactions are mostly unclear. In this study, we characterized a novel extracellular β‐galactosidase (Amuc_1686) with a preference for linkages from the type Galβ1–3GalNAc. Additionally, Amuc_1686 possesses a discoidin‐like domain, which enables the interaction with anionic phospholipids. We detected a strong inhibition by phosphatidylserine, phosphatidylglycerol, phosphatidic acid, and lysophosphatidic acid while phosphatidylcholine and phosphatidylethanolamine had no influence. Amuc_1686 is the first example of a prokaryotic hydrolase that is strongly inhibited by certain phospholipids. These inhibiting phospholipids have important signal functions in immune response and cell clearance processes. Hence, Amuc_1686 might be regulated based on the health status of the large intestine and could therefore contribute to the mutualistic relationship between the microbe and the host on a molecular level. In this sense, Amuc_1686 could act as an altruistic enzyme that does not attack the mucin layer of apoptotic epithelial cells to ensure tissue regeneration, for example, in areas with inflammatory damages.The gut microbe  Oligonucleotides were synthesized by Eurofins .All chemicals, reagents and substrates used in this study were purchased from Carl Roth GmbH , Sigma‐Aldrich , or Megazyme . 2.2Escherichia coliDH5α and E. coliBL21 (DE3) were grown in lysogeny broth  harboring plasmids of interest were used to inoculate 1 L LB medium and were incubated at 37°C in shaker flasks at 180 rpm. When the cultures reached an optical density at 600 nm of approximately 0.4, protein production was induced by addition of 0.2 µg ml‐1 anhydrotetracycline. Cells were harvested at an optical density between 1.0 and 1.5 by centrifugation at 9,000 g. Lysis and purification was done as previously described  at optimal pH (7.5) and temperature (65°C). Inhibition experiments were performed at 37°C and a 50 mM combined buffer system pH 7.5 was used. All potentially inhibiting compounds were dissolved in methanol prior to assay application. The impact of different ions on the enzyme activity was analyzed in 100 mM Tris‐HCl buffer pH 7.5.Enzyme activity assays for Amuc_1686 were performed in a combined buffer system containing sodium acetate, Tris‐HCl, potassium dihydrogen phosphate and dipotassium phosphate (50 mM each). In case of colorimetric 2.62SO4as mobile phase at 25°C and a flow rate of 0.6 ml min‐1. The amounts of products and the grade of substrate degradation was determined, using an UV‐Vis detector at 210 nm and a refractive index detector. Products were quantified by comparison to calibration curves.For substrate spectrum analysis with non‐colorimetric substrates, enzymatic assays were performed as described above and incubated for different time periods at 37°C. Samples were applied to HPLC using an Aminex‐HPX87H column  with 5 mM H33.1A. muciniphila ATCC BAA‐835. The corresponding gene amuc_1686encodes a protein with a molecular mass of 86 kDa. BLAST analysis of Amuc_1686 using the SwissProt database indicated that 19 of the 20 closest hits were exclusively eukaryotic β‐galactosidases from mainly mammalian species such as Homo sapiens(Q8IW92), Mus musculus(Q3UPY5), and Rattus norvegicus(Q5XIL5). Only one protein with significant homology  was found in the bacterial kingdom, the β‐galactosidase Bga from Xanthomonas manihotis(P48982) .Gel filtration chromatography revealed a monomeric native structure for Amuc_1686. Purified Amuc_1686 showed the highest catalytic efficiency at 65–70°C and at pH 7.5. Substrate spectrum analysis was performed using various artificial chromogenic nitrophenyl‐linked sugars and natural di‐ and trisaccharides Table  at 65°C N‐biose, consisting of galactose that is β‐1,3 glycosidic bound to N‐acetyl‐d‐galactosamine . Examination of product formation revealed a cleavage of approximately 98% galacto‐N‐biose within 6 hr with an activity of 7.6 ± 0.5 U mg‐1 , a N‐acetyl‐beta‐hexosaminidase from Clostridium perfringensATCC 13124 (NagJ), and a hyaluronoglucosaminidase from C. perfringens (NagH) , phosphatidylglycerol (PG), phosphatidic acid (PA), phosphatidylcholine (PC) or phosphatidylethanolamine (PE) dissolved in methanol were added to the assay and the inhibiting effect of the phospholipid was analyzed. PC , 5.37 (PA) and 6 (PG). Additionally, the potential inhibiting influence of lysophosphatidic acid (LPA) was investigated, showing an IC50 value of 13.6 ± 10.6 µmol L‐1 which corresponds to a pIC50 value of 4.87 (not shown). Control experiments using pure methanol did not reveal a negative influence on the enzymatic activity.It is known that discoidin domains, for example, found in the carboxylterminus of blood coagulation factors 5 and 8, can promote binding to cell surface phospholipids such as phosphatidylserine and are responsible for enzyme activity , lacking the last 107 amino acids. For inhibition experiments, 1 µg of Amuc_1686_short was applied to a standard assay that was identically performed as with Amuc_1686. When directly compared with Amuc_1686, the shortened variant exhibited a slightly decreased enzymatic activity but showed no significant inhibitory effect after addition of PS, PG and PA in the same concentration as applied to the assay containing the full‐length enzyme Figure .l‐serine, glycerol, choline and ethanolamine were added to the standard assay up to a final concentration of 3 mmol L‐1. Under these conditions, no inhibition was observed. To identify the type of inhibition, the Km value for para‐nitrophenyl‐β‐d‐galactopyranoside in the presence of 0.25 µmol L‐1 PG was determined. As the measured values were in the same range as without inhibitor addition, we concluded a non‐competitive type of inhibitory effect of the phospholipids on Amuc_1686.In order to analyze whether the hydrophilic head groups of the phospholipids have an influence on enzyme activity, 4A. muciniphila is able to utilize the mucus layer that covers colonic epithelial cells in the human large intestine. Mucin is used by this organism as carbon and nitrogen source which has an ecological advantage due to limitation of free carbon sources in this specific gut region GalNAc‐O‐S/T] mucins. In the human transversal and descending colon, the natural habitat of A. muciniphila, the major part of mucin oligosaccharide structures is based on core structure type 3 [GlcNAcβ1–3GalNAc‐O‐S/T] or type 4 [GlcNAc1–3(GlcNAcβ1–6)GalNAc‐O‐S/T] GlcNAcβ1–3GalNAc‐O‐S/T] (Robbe et al., The only hydrolytic activity of Amuc_1686 with a non‐chromogenic substrate was detected with Galβ1–3GalNAc (Table A. muciniphila by the anionic phospholipids PA, PG, PS and LPA. It became evident that the C‐terminal discoidin domain of the enzyme is responsible for this regulatory effect because the shortened variant of Amuc_1686, missing the discoidin domain, showed no significant inhibition by these lipids.Analysis of the primary structure of Amuc_1686 revealed a unique C‐terminal domain which can be bioinformatically classified as discoidin domain Figure . In euka50of 15 nM (Jones & Hannun, 2+‐ATPase of the sarcoplasmic reticulum (Dalton et al., A. muciniphila und the human intestinal tract, this regulatory system might reflect a mechanism which allows this mutualistic relationship (Figure In eukaryotic cells, enzyme inhibition by phospholipids was previously described (Stace & Ktistakis, A. muciniphilapositively influences and promotes wound healing and tissue regeneration (Alam et al., A. muciniphila and its host, a regulation of the extracellular mucin‐degrading enzyme Amuc_1686 on protein level is conceivable (Figure It is known that e Figure . An inteIn addition to PS, a strong inhibitory regulation of Amuc_1686 was observed by the anionic phospholipids PA, PG and LPA. PA is known to serve as a lipid second messenger (McPhail et al., Another aspect is that carbohydrate chains of the cellular surface function as a signal for apoptotic cell removal (Beppu, Eda, Fujimaki, Hishiyama, & Kikugawa, A further example for a mucin‐like structure involved in immune response is the glycoprotein lactadherin which recognizes and binds to PS by a discoidin domain. This protein contains mucin‐like glycosidic structures, enhances phagocytosis and is crucial for repair of intestinal epithelium (Bu et al., A. muciniphila is limited from damaged areas of the large intestine. However, further experiments have to be done in the future to verify or to refute the proposed hypothesis.In summary, Amuc_1686 can be classified as a phospholipid‐regulated enzyme that might not attack apoptotic epithelial cells to ensure tissue regeneration and to avoid a disorder of the clearance process. This mutualistic behavior would support health of the host but also implicates that substrate for growth of There is no conflict of interest for all authors.Uwe Deppenmeier and Konrad Kosciow conceived and designed the research. Konrad Kosciow performed the experiments and wrote the initial manuscript draft. Uwe Deppenmeier supervised the research and revised the manuscript draft.None required.


Scanning tunneling microscopy resolves characteristic charge orders within domain walls and their vortices. Density functional theory calculations disclose their unique atomic relaxations and the metallic in-gap states confined tightly therein. A generic theory is constructed, which connects this emergent honeycomb network of conducting electrons to the enhanced superconductivity.When two periodic potentials compete in materials, one may adopt the other, which straightforwardly generates topological defects. Of particular interest are domain walls in charge-, dipole-, and spin-ordered systems, which govern macroscopic properties and important functionality. However, detailed atomic and electronic structures of domain walls have often been uncertain and the microscopic mechanism of their functionality has been elusive. Here, we clarify the complete atomic and electronic structures of the domain wall network, a honeycomb network connected by Z 2, which may help understand the enhanced superconductivity.The detailed structures of domain walls are important to determine macroscopic properties in charge density wave materials. Here, Park et al. report atomic and electronic structures of a honeycomb domain wall network in 1T-TaS For example, the commensuration interaction in graphene layers induces various superstructures of discommensuration with exotic electronic property such as unconventional superconductivity10. Discommensurations are also popular and important in 2D systems with more complex interactions, such as charge ordered or charge-density wave (CDW) systems, where domain wall networks have been suggested crucial in metal–insulator transitions12, emerging superconductivity15 and the formation of various metastable states for device functionality18.Competing periodicities and discommensuration phenomena have been widely discussed from one-dimensional (1D) chain models2 CDW are outstanding21, as combined further with the strong electron correlation22 and the strong spin frustration24. 1T-TaS2 is composed of layers of hexagonal tantalum atoms coordinated octahedrally by sulfur atoms. A metallic phase at high temperature transits into an incommensurate CDW phase below 550 K, a nearly commensurate CDW (NC-CDW) below 350 K, and a commensurate CDW (C-CDW) below 180 K11. The atomic structure of the C-CDW phase has the unit of the well-known David-star cluster; among 13 Ta atoms of a cluster 12 are distorted toward the central one forming a 25. One unpaired electron in the central Ta atom falls into the Mott-insulator state by onsite Coulomb repulsion22. The NC-CDW phase was reported to consist of C-CDW domains and a honeycomb domain-wall network31 with its atomic structure still unknown and its electronic property under debates16. Recent experiments reported that various metallic excited states are formed, presumably with highly conducting domain wall networks, by pressure, doping and other control parameters37. These states may be utilized in a ultrafast memory16 or a memristor device17. The domain walls are also suggested as the origin of superconductivity emerging from those metallic states39. However, recent STM experiments observed the nonmetallic nature of domain walls for the voltage-pulse-induced domain-wall phase34 and isolated domain walls at low temperature40. Under these contradictory findings, the characterization of atomic and electronic properties of domain walls in the NC-CDW phase and the general microscopic understanding of the role of the domain wall network for emerging quantum phases and functionalities are highly requested.Among diverse 2D materials, exotic low-dimensional electronic properties and domain walls of 1T-TaS2 by scanning tunneling microscopy (STM). By comparison with the experiment, our density functional theory (DFT) calculations identify that the domain wall consists of particular types of imperfect David-star clusters. Domain walls are further connected by Z3 topological vortices of domain walls to form a honeycomb network. We also identify the atomic structure and the novel charge order within these vortex cores. The metallic states of the NC-CDW phase originate from imperfect David stars of domain walls and vortices. A generic theory for the conducting honeycomb network indicates symmetry-protected Dirac bands, quadratic band touchings, and flat bands, which would definitely play important roles in the emerging superconductivity. This work provides unprecedented microscopic understanding of discommensurations in 2D materials with strong interactions, which has wide implication into diverse 2D materials with open possibilities for designing functionalities and quantum phases through the discommensuration engineering.In this work, we directly resolve atomistic details of discommensuration domain walls of the NC-CDW phase in 1T-TaS2 at room temperature. It consists of the domains with relatively high CDW amplitude separated by the low-amplitude regions of domain walls. A hexagonal array of domains and CDW maxima within each domain are more clearly seen in the low-pass filtered image of Fig. qCDW = 0.519 Å−1 (12.11 Å) in consistency with the periodicity of C-CDW21. In addition, the main peaks carry satellites42 rotated slightly with respect to the C-CDW vector (see the enlargement)44, which have qHEX = 0.083 Å−1 (76.13 Å) with a rotation of 9.5 ± 2° from the C-CDW peaks. This represents the hexagonal lattice of domains and a honeycomb domain-wall network.Figure b (a × b is the inplane unitcell of the undistorted structure) among the 12 possible configurations due to 13 Ta atoms within one C-CDW unitcell as classified previously34. This particular configuration is found ubiquitously along all hexagonal directions between domain centers over the whole area probed. In the domain area, a single David star exhibits mainly three bright spots arranged in an inverted triangle. They correspond to top three S atoms at the center of a David star cluster. In contrast, major atomic protrusions in the domain wall are arranged mostly in regular triangles  with about 19 perfect David-stars for each domain and with the angles with respect to the C-CDW vector of 9.6°. These structural parameters agree well with the present and the previous experiments26. The center position of David-star clusters in each domain is shifted28 and domains are connected with the DW-1 structure, which is the key atomic structure of the NC-CDW phase. Note also that this structure has regions where three domain walls meet  can be defined to have a clockwise (anticlockwise) rotation of phase-shift displacement vectors between involved domains  lattice compression up to about 5 (6.7)%. This means that the Mott state of the domain regions of NC-CDW phase would also be robust. On the other hand, the interlayer interaction would affect the electronic structure of the NC-CDW phase by the out-of-plane band dispersion and the change of the in-plane electronic structure by a possible stacking order49. These recent suggestions indicate metallic or small-gap insulating states, which are not consistent with the large Mott gap observed and are fragile against electron correlation /L)   applied to the sample. A single crystal of 1T-TaS2 was prepared by iodine vapor transport method. The samples were cleaved at room temperature and quickly transferred to the precooled STM head.The STM measurements have been carried out with a commercial microscope (Omicron) at room-temperature. Pt–Ir wires were used as STM tips and topographic images were acquired in the constant current mode with bias voltage (61 within the Perdew–Burke–Ernzerhof generalized gradient approximation62 and the projector-augmented wave method63. The single-layer 1T-TaS2 was modeled with a vacuum spacing of about 13.6 Å. We used a plane-wave basis set of 259 eV and a 6 × 6 × 1 k-point mesh for the d orbitals, which was estimated as 2.27 eV by the linear-response method64. This value reproduces the experimental Mott gap size21. To obtain the band spectrum of the honeycomb network model, we generalized the network model51 on a triangular lattice to that on the honeycomb lattice. It is equivalent to the honeycomb array of scatterers of 1D metals and we solved it by using the standard Bloch wavefunction.DFT calculations were performed by using the Vienna ab initio simulation packageSupplementary InformationPeer Review File


Plasmodium falciparum (Pf) and P. vivax (Pv) are the most virulent and responsible for the majority of the mortality and morbidity attributed to malaria.Malaria affects one third of the world’s population and kills hundreds of thousands across the globe annually. Among the species that cause malaria in humans, Anopheles mosquito bites an individual and releases Plasmodium sporozoites, the motile and infective form of the parasite, into the skin. The deposited sporozoites migrate through the skin, enter the circulatory system, and traffic to the liver. This migration requires traversal of the sporozoite through diverse cell types of distinct host tissues. Inside the liver, Plasmodium sporozoites first undergo mandatory, asymptomatic, acyclic intrahepatic development before mature merozoites egress and infect red blood cells. The cyclic blood stage involves repeated rounds of invasion and replication within red blood cells and is responsible for all the disease symptoms and complications. During the blood stage, some parasites develop into gametocytes, which are the first step in the sexual cycle. The Plasmodium life cycle continues as a mosquito feeds on an infected host and takes up gametocytes in a blood meal. The parasites undergo sexual reproduction in the mosquito, which results in mature sporozoites that then propagate the next cycle of infection.Malaria infection begins when an infected female Plasmodium is predominantly intracellular during blood stage growth, and this protects parasites from the host immune response. However, the parasite is vulnerable when it is extracellular during traversal and prior to host cell invasion. Components of the Plasmodium invasion machinery are pathogen-specific and surface-exposed, making them potential vaccine and/or drug targets that can be exploited to design therapeutics against the deadly parasite. Structural studies provide the precise definition of the linear and nonlinear conformational neutralizing epitopes that can be exploited to improve the immunogen design of malaria vaccines. In addition, the structural details of Plasmodium specific ligands and ligand/receptor complexes provide unprecedented insight into the mechanisms of interaction, invasion, and inhibition at the host–parasite interface.Plasmodium-specific invasion proteins involved in 1) traversal, 2) hepatocyte invasion, and 3) erythrocyte invasion . The current review focusses on the most virulent human malaria parasites, P. falciparum and P. vivax. Additional components of the invasion machinery are also excluded from the current study as they have been extensively reviewed elsewhere.In this review, we describe the structural and functional details of Plasmodium uses stage-specific pore-forming proteins to disrupt host cell membranes to either enter or exit host cells during traversal, and to egress from the parasite-built parasitophorus membrane after invasion and replication. The characterized pore-forming proteins include sporozoite protein essential for cell traversal 1 (SPECT1), perforin-like proteins (PLPs), and cell traversal protein for ookinetes and sporozoites (CelTOS) .Sporozoites are deposited into the dermis of the host upon the bite of an infected mosquito. These motile sporozoites traverse through the skin to find blood vessels and subsequently reach the liver through the circulatory system. Before a sporozoite can invade and replicate in a hepatocyte, it must traverse several physical barriers, including fibroblasts, Kupffer cells, and sinusoidal endothelial cells, to reach the target hepatocyte. Host cell traversal is the process of parasite entry into, passage through, and egress from host cells without lysis. Host cell traversal protects the vulnerable sporozoite from phagocytosis, primes the sporozoite through the activation of apical exocytosis, and prepares the motile sporozoite for invasion . In addiPlasmodium proteins that may have possible roles in cell traversal. The targeted disruption of P. falciparum SPECT1 or PLP1 cause reduced infectivity of sporozoites in liver-stage development in humanized mice [SPECT1 and PLP1 are essential zed mice . HoweverPlasmodium pore-forming protein that is required for cell traversal in both the mammalian host and the mosquito vector. Recently, the crystal structure of P. vivax CelTOS revealed an all α-helical, tuning fork–shaped dimer structure that resembled membrane-disrupting proteins from viruses and bacteria  [P. vivax and P. falciparum CelTOS can bind phosphatidic acid (PA), an inner leaflet abundant lipid, suggesting a plausible inside-out function of this protein during parasite traversal . CelTOS forms pores in liposomes containing PA as observed by negative stain transmission electron microscopy. The protected hydrophobic core of this soluble dimer CelTOS structure suggests that a significant conformational rearrangement is mandatory to form a pore in cell membranes. Further work is needed to explore the conformation of the lipid-bound CelTOS. Antibodies raised against P. falciparum CelTOS protect from infection, and thus, CelTOS is a vaccine candidate being evaluated currently in clinical trials [Cell traversal protein for ookinetes and sporozoites (CelTOS) is a unique  . P. vival trials .circumsporozoite protein (CSP), thrombospondin-related adhesive protein (TRAP), and P36 .Following initial cell traversal, migratory sporozoites establish hepatocyte infection and undergo exponential growth to develop tens of thousands of primary merozoites. During active invasion, sporozoite surface proteins interact with host receptors to facilitate entry into the host cell. The three most actively studied sporozoite surface coat proteins are 25-49) which is diverse among different Plasmodium species. This repeat region is flanked by conserved N- and C-terminal domains. The N-terminus of CSP contains a charged protease-cleavage site known as region I (RI). The C-terminus contains a short, conserved sequence in region III (RIII) and a known thrombospondin-like type 1 repeat (TSR1) cell adhesive motif . Antibodies against PfCSP C-terminal, induced by live sporozoite vaccination in humans, are ineffective against the malaria infection [CSP is the most abundant surface protein on sporozoites and has multiple roles in sporozoite development, gliding motility, and active invasion. CSP is composed of a central repeat region  [P. falciparum CSP revealed a unique αTSR domain [Plasmodium species  and αTSR domain are components of the leading malaria vaccine RTS,S/AS01 [18), highly neutralizing anti-CSP antibodies engage the junctional residues between the N-terminal domain and repeat region [The individual CSP domains function in a stage- and time-specific manner. RI recognizes heparan sulfate proteoglycans (HSPG) on the salivary gland in the mosquito vector , and thiFig 1B) . The cryR domain , where tS,S/AS01 . RTS,S/AS,S/AS01 –8. In ad [ [Plasmodium, stage-specific TRAP members exist that are essential for the invasion of the merozoite (MTRAP), ookinete (CTRAP), or salivary gland sporozoites (TRAP) [P. falciparum TRAP using a systematic extracellular protein screening approach [TRAP also contains a TSR domain and is involved in host invasion and gliding motility. TRAP localizes to the plasma membrane and translocates from the anterior to the posterior end of the sporozoite during invasion. In TRAP, the N-terminus encompasses a rigid domain that shows structural homology to the von Willebrand factor (vWF) type A-domain ( . This vW . When a  [ . In Plass (TRAP) . Recentlapproach . An inacapproach .P. falciparum binds the host receptor CD81, while P. vivax P36 binds to scavenger receptor BI (SR-B1) as the hepatocyte receptor during effective infection [Plasmodium parasites contain fourteen 6-cysteine domains containing proteins, which are conserved among different Plasmodium species and play crucial role in fertilization, parasitophorous vacuole membrane fitness, and immune evasion [P36 is a 6-cysteine domain containing protein. Recent genetic analyses proposed a role for P36 in liver-stage invasion , 18. Int evasion . Therefo evasion .P. falciparum invading reticulocytes as well as mature erythrocytes, while P. vivax is specific for reticulocytes.Primary merozoites released from ruptured hepatocytes enter into the blood stream, invade erythrocytes, and develop into ring, trophozoite, and schizont stages, culminating in the formation of 16 to 32 mature merozoites. Each of these merozoites can invade a fresh erythrocyte and continue the cyclic, asexual blood stage development. Malaria parasites also exhibit distinct red cell tropism with Fig 1C). Reticulocyte-binding ligand (RBL) and erythrocyte-binding like (EBL) are two critical protein families involved in red blood cell invasion .Unlike all other stages, the fluidic nature of blood stage infection subsides the requirement of early traversal but complicates the invasion process. Red cell invasion by the parasite involves 1) initial interactions causing erythrocyte deformation, 2) apical interactions and invasion, and 3) a final recovery phase. This review covers the apical invasion and the parasite ligands involved therein  is the reticulocyte binding family that consists of PfRh1, PfRh2a, PfRh2b, PfRh4, and PfRh5. Receptors have been identified for PfRh4 (complement receptor 1) [eptor 1)  and PfRheptor 1) . While aeptor 1) .P. falciparum [Fig 1C) [PfRh5 is a leading blood stage vaccine candidate and an exceptional member of the Rh family as antibodies that prevent the interaction of PfRH5 with the host receptor basigin neutralize diverse lab and field isolates of lciparum . The PfR[Fig 1C) . Rh5 for[Fig 1C) , 26. Alt[Fig 1C) . Rh5 lac[Fig 1C) , 28. Div[Fig 1C)  and memb[Fig 1C) , howeverP. vivax reticulocyte-binding proteins (PvRBPs) are the homologs of PfRhs, and this family is composed of 11 proteins [proteins .P. vivax, although maturation of DARC has also been proposed [P. vivax RBP2b:TfR1:Tf ternary complex reveals how P. vivax uses PvRBP2b to hijack host TfR1 to invade the host reticulocyte without affecting binding of TfR1 to Tf [Fig 1C). Monoclonal antibodies raised against PvRBP2b prevent reticulocyte binding and reduce P. vivax invasion [PvRBP2b was recently shown to bind transferrin receptor 1 (TfR1), a highly expressed surface receptor on a variety of mammalian tissues, including reticulocytes, that delivers iron-loaded transferrin (Tf) glycoprotein into cells to maintain iron homeostasis . TfR1 isproposed . A recenR1 to Tf . PvRBP2binvasion .P. vivax and consists of a conserved kite-shaped domain but possesses distinct surface properties, suggesting a recognition site for the alternate receptor  [PvRBP2a is another structural homolog of PfRh5 in  . In contPlasmodium-specific, host receptor–binding domain, region II (RII). In EBL-proteins, RII contains either a single or double Duffy Binding–like (DBL) domain. EBL-family proteins interact with the host receptor through the RII region. In P. falciparum, four EBL-proteins have been identified: PfEBA-175, PfEBA-140, PfEBA-181, and PfEBL-1. P. vivax has a single member known as Duffy binding protein (PvDBP).EBL-family proteins are a second set of redundant invasion ligands that contain a conserved domain architecture including a conserved, Fig 1C) [PfEBA-175 interacts with glycophorin A (GpA) on the erythrocyte surface in a sialic acid-dependent manner. The crystal structure of RII in complex with ⍺-2,3-sialyllactose revealed that RII is a dimer and that the sialic acid binding sites are located at the dimer interface  . Site-diFig 1C) , suggestFig 1C) . The proFig 1C) , 37. A PFig 1C) . EBA-175Fig 1C) .P. falciparum DBL-domain–containing protein that binds to glycophorin C (GpC) in a sialic-acid–dependent manner. The structure of PfEBA-140 RII domain, consisting of two DBL domains, revealed two sialic acid binding sites within a monomer of RII, each of which were contained within its respective DBL domain  [P. falciparum lab strains, suggesting the role of this ligand in parasite invasion, with antibodies that targeting F1 having a greater neutralizing potential [PfEBA-140 is another otential , 42.PfEBA-181 and PfEBL-1 are two other DBL domains containing proteins that have conserved EBL-family architecture but are not yet structurally characterized.P. vivax. PvDBP binds the Duffy antigen receptor for chemokines (DARC) on the reticulocyte surface. Unlike PfEBA-175, the RII domain of PvDBP consists of a single DBL domain . In vitro structural and biophysical studies of PvDBP RII identified a conserved stoichiometry of 2:2 between PvDBP and the host receptor DARC [P. vivax DBP-DARC interface to reduce malaria. Although PvDBP is polymorphic, broadly neutralizing epitopes have been identified in the DBL domain [P. vivax tropism [PvDBP is the functional ortholog of PfEBA175 in tor DARC . The sultor DARC . The strtor DARC  is distitor DARC . The rector DARC . TherefoL domain . Interes tropism .Plasmodium invasion machinery and its individual components in the last few decades. Future research will leverage the available structural information and explore the conformational space of the invasion machinery to design, develop, and optimize novel therapeutics. The preliminary success of RTS,S/AS01, the first marketed malaria vaccine, supports the idea that an infection-blocking malaria vaccine is indeed feasible. Understanding the mechanisms of Plasmodium invasion will guide development of novel vaccines to interrupt the invasion process and prevent disease and transmission of malaria.In conclusion, through advanced tools in genetics, structure biology, and immunoparasitology, we have gained immense knowledge about the


Three-dimensional modeling of the HMA4 protein in Arabidopsis reveals the zinc permeation pathway across the plasma membrane, and mutations in the pathway alter zinc and cadmium transport differently in plants. 1B ATPase heavy metal ATPase 4 (HMA4) is responsible for zinc and cadmium translocation from roots to shoots in Arabidopsis thaliana. It couples ATP hydrolysis to cytosolic domain movements, enabling metal transport across the membrane. The detailed mechanism of metal permeation by HMA4 through the membrane remains elusive. Here, homology modeling of the HMA4 transmembrane region was conducted based on the crystal structure of a ZntA bacterial homolog. The analysis highlighted amino acids forming a metal permeation pathway, whose importance was subsequently investigated functionally through mutagenesis and complementation experiments in plants. Although the zinc pathway displayed overall conservation among the two proteins, significant differences were observed, especially in the entrance area with altered electronegativity and the presence of a ionic interaction/hydrogen bond network. The analysis also newly identified amino acids whose mutation results in total or partial loss of the protein function. In addition, comparison of zinc and cadmium accumulation in shoots of A. thaliana complemented lines revealed a number of HMA4 mutants exhibiting different abilities in zinc and cadmium translocation. These observations could be instrumental to design low cadmium-accumulating crops, hence decreasing human cadmium exposure.The P IB ATPases, a subfamily of P-type ATPases, are metal cation transporters essential for the survival of organisms ranging from bacteria to eukaryotes such as plants and animals  transporters. This subgroup is found in all domains of life and includes the well-characterized CopA bacterial transporter, as well as the human ATP7A and ATP7B proteins, whose mutations lead, respectively, to Menkes syndrome and Wilson’s disease  and are only present in bacteria and plants, with, for instance, the bacterial pumps ZntA and CadA and the Arabidopsis thaliana heavy metal ATPases 2–4 (HMA2–HMA4)  forming the TM domain, two cytoplasmic catalytic domains, the actuator domain (A domain), and the ATP-binding domain (ATP domain) divided into a nucleotide-binding domain (N domain) and a phosphorylation domain (P domain), and for most of them N- and occasionally C-terminal cytosolic extensions. As members of the P-type ATPase superfamily, these proteins use the energy of ATP to transport their substrate following the E1/E2 Post–Albers cycle  with a typical βαββαβ-fold structure containing a consensus CxxC motif binding one Cu+ or one Zn2+ ion, respectively  preceded by two additional helices in many of them (TMA and TMB). A characteristic sequence of three amino acids (CPx motif) in TM4 together with amino acids in TM5 and TM6 may determine ion selectivity 7K and DxG(x)7N on TM5 and TM6, respectively (i) of the Shigella sonnei ZntA protein, a homolog of HMA4, has recently been determined  that directs free Zn2+ ions to the intramembrane Zn2+-binding site. This site consists of C392 and C394 of the conserved CPx motif and D714 of the DxG(x)7N PIB-2 signature sequence. Their involvement in Zn2+ binding had already been characterized biochemically 7K motif] located near D714 forms a salt bridge with the latter, preventing Zn2+ reflux to the binding site  (4 in a short-day growth chamber (22 °C and 8 h d–1 photoperiod) during 7 weeks. They were then transferred in long days (16 h d–1 photoperiod) where they were supplied with 3 mM ZnSO4 for 5 weeks to allow flowering. The plants were transformed using Agrobacterium tumefaciens by floral dipping ( dipping .1) heterozygous transgenic seeds  were germinated on hygromycin B (20 µg ml–1) 1/2 MS (Murashige and Skoog) agar medium containing 1% sucrose in short days. After 14 d, seedlings were transferred to soil , watered with tap water, and grown for 5 weeks in long days prior to imaging and sample harvesting.For phenotyping on soil and metal accumulation analysis, first-generation  fragment was cloned into the pAhHMA4-2::AhHMA4::GFP pBluescript II KS+ vector . Site-directed mutagenesis, using the QuickChange Site-Directed Mutagenesis method (Agilent Technologies) and mutagenic primers (see JXB online), was then performed on the newly constructed vector to create the TM variants  on a DigiPrep Graphite Block Digestion System (SCP Science) as follows: 15 min at 45 °C, 15 min at 65 °C, and 90 min at 105 °C. After cooling, sample volumes were adjusted to 10 ml with milliQ water, and 200 µl of ≥65% HNO3 was added. Metal concentrations were determined using inductively coupled plasma atomic emission spectroscopy (ICP-AES) with a Vista-AX instrument  as described (Shoot tissues were cleaned with milliQ water and dried at 60 °C for 3 d. Shoot samples (10–50 mg of tissues) were then acid-digested in DigiPrep tubes with 3 ml of ≥65% HNO1 seeds of A. thaliana hma2hma4 plants expressing the WT and variant AtHMA4 proteins fused to green fluorescent protein (GFP) as described above were germinated on hygromycin B (20 µg ml–1) 1/2 MS agar medium containing 1% sucrose in short days. After 14 d, seedlings were transferred on the same medium without antibiotic. After 3 d, roots of 2–4 independent lines per construct from two independent experiments were analyzed. Images were collected at a 1024 × 1024 pixel resolution using a TCS SP5 inverted confocal laser microscope (Leica Microsystems) with a ×63 water immersion PlanApochromat 1.20 objective (Leica Microsystems) as previously described (https://imagej.nih.gov/ij/) and plot profile analysis. Briefly, in pericycle cells expressing HMA4 fused to GFP, 10 optical sections were drawn across the transversal plasma membranes. GFP fluorescence intensity values (n=20) were then used to calculate a mean fluorescence intensity for each independent mutant line.Thttp://www.uniprot.org). This protein has an N-terminal MBD (residues 1–96) that has been resolved by NMR (PDB ID: 2KKH) (https://blast.ncbi.nlm.nih.gov) (http://www.rcsb.org/pdb)  (https://swissmodel.expasy.org) through the QMEAN6 Z-score (The Uniprot code of the AtHMA4 protein is O64474 (sn-glycero-3-phosphocholine) bilayer of 347 lipids with the insane tool (XY and Z. Non-bonded interactions were computed up to 1.2 nm with the shift method. Electrostatics were treated with dielectric permittivity constant ε=15. The compressibility was 10–5 (1/bar). The system was then transformed to an atomistic resolution with backwards (–5 (1/bar) and τP=5 ps. Non-bonded interactions were evaluated using a twin-range cut-off scheme. Interactions within the shorter range cut-off (0.8 nm) were calculated every step, whereas interactions within the longer cut-off (1.4 nm) were updated every five steps, together with the pair list. In all the simulations, a reaction ﬁeld correction was applied to the electrostatic interactions beyond the long-range cut-off of 35 using a relative dielectric permittivity constant εRF of 62 (http://www.PyMOL.org) and VMD software  in HMA4 instead of one in ZntA (G391) . Additioin planta the functional importance of the amino acids depicted in the model, HMA4 transmembrane variants were generated by directed mutagenesis (HMA4 (WT) and mutated genes were fused to the GFP gene (HMA4::GFP) and expressed under the control of the endogenous A. halleri HMA4 promoter 2 (pAhHMA4-2) in the loss-of-function hma2hma4 A. thaliana mutant  accumulated ~5-fold higher zinc in shoots than the hma2hma4 mutant, reflecting the inability of the latter to translocate zinc from root to shoot   bordering the motif  was significantly decreased, suggesting that HMA4 might be more Zn efficient than many of its plant homologs  and Oryza sativa HMA2 (OsHMA2) and HMA3 (OsHMA3) proteins  for molecular dynamic simulations.Fig. S1. Amino acid sequence alignment of Shigella sonnei ZntA and Arabidopsis thaliana HMA4.Fig. S2. Complementation of the A. thaliana hma2hma4 zinc deficiency phenotype.Fig. S3. HMA4 TM variant localization in A. thaliana.Fig. S4. HMA4 TM variant expression level in A. thaliana.Fig. S5. Sequence conservation of the TM region among plant PIB-2 ATPases.Fig. S6. Distribution of residue types at the surface of the N-terminal MBDs of HMA4 and ZntA.Fig. S7. View of the transmembrane domain of HMA4 in the E2P and E2-Pi states in a membrane environment during molecular dynamic simulations.Fig. S8. Amino acid sequence alignment of Arabidopsis thaliana HMA4, Nicotiana tabacum HMA4, Oryza sativa HMA2, and Oryza sativa HMA3.Supplementary Figures S1-S8 and Tables S1-S2Click here for additional data file.


The destructive nature of subduction erosion poses challenges to fully understanding the evolution of erosive convergent margins that are critical to understanding crustal recycling and seismogenesis. Forearc deformation holds important clues to the evolution of erosive convergent margins. Here we present detailed paleomagnetic and structural analyses of IODP Site U1380 cores from the middle slope of the forearc of the Costa Rican erosive convergent margin. The analyses reveal a strong deformation zone from ~490 to ~550 mbsf that is characterized by abundant fissility/foliations shallower than the bedding. Similar relatively strong deformation zones are recognized from the frontal prism and upper slope sites, and are broadly correlative, forming a zone of strong deformation across the forearc. This zone spans ~2.0 to 1.83 Ma and the deformation likely occurred briefly at ~1.80 Ma. The widespread, short-lived, and strong deformation is interpreted as a result of intense subhorizontal shear following the rapid forearc subsidence driven by the dramatic subduction erosion associated with the abrupt onset of the Cocos Ridge subduction. Given the typical occurrence of forearc subsidence by subduction erosion, similar styles of deformation are probably common in other erosive convergent margins as well. About half of the worldwide convergent margins are erosive3. Erosive convergent margins are characterized by subduction erosion at the frontal and/or basal part of forearc wedges7 and thus important for crustal material transfer. Erosive convergent margins are also the places where large earthquakes occur8. Therefore, elucidating the evolution of erosive convergent margins are critical in understanding crustal recycling9 and origin of earthquakes10 in subduction zones. However, the destructive nature of subduction erosion poses significant challenges in deciphering the rock record at erosive convergent margins. Nevertheless, forearc wedges in erosive convergent margins are situated at the forefront in the subduction process and their deformation histories hold important clues to the evolution of this type of convergent margin.Convergent plate boundaries are considered either accretional or erosive on the basis of whether material is accreted onto or eroded away from the overriding plate in subduction zones11  at ~9.0 cm/yr20 at a shallow angle (<10°)21 beneath the Caribbean plate  Expeditions 334 and 344 cored not only the frontal prism (Site U1412), but also the subducting CR (Sites U1381 and U1414) and middle slope (Sites U1378 and U1380), and the upper slope (Sites U1379 and U1413) offshore southern Costa Rica (near Osa Peninsula)29 , an overthickened aseismic ridge 22 Fig. . The CR 29. In addition, a detailed paleomagnetic study refines the chronological framework and constrains the deformation history at the middle slope site. Furthermore, a comparative analysis of the deformation in the middle slope, the frontal prism, and the upper slope allows an improved understanding of the forearc deformation of the Costa Rican erosive convergent margin.In this paper, we examine the deformation style and history at the middle slope Site U1380 in the southern segment of the Costa Rican margin. The deformation style is investigated using the anisotropy of magnetic susceptibility (AMS) together with the structural data collected during the expeditions33 , clayey siltstone and sandstone (Unit II), and silty clay (Unit I). Samples from all three units show predominantly oblate fabrics33 Fig. . While at I Fig. . For Unit I Fig. .Figure 2min perpendicular to the horizontal bedding as gravity is the predominant force34. The Kmin tilt off the vertical could be depositional or tectonic related (tilted beds) in origin. Sediment deposition on a slope, or in the presence of current35, or grain collision or viscous suspension during sediment settling36 can lead to Kmin tilt off the vertical. Kmin tilt of the oblate AMS fabrics of the deep sea sediments from the Nankai Trough accretionary prism has been employed to investigate the sediment transport/deposition processes37 and the degree of deformation in the Nankai Trough accretionary prism38. Similarly, the Kmin tilt of AMS fabrics in the upper hemipelagic/pelagic sediments on the subducting Cocos plate of the Costa Rican convergent margin is believed to be associated with the initial underplating of sediments beneath the margin15. However, the Kmin tilt at the base of the hemipelagic unit was likely related to the tilted strata because steep bedding dip was observed14.It is well known that deposition of sediments in still water can produce oblate fabrics that are characterized by K8. The dip of the inclined bedding was measured and documented by the shipboard structural geologists during the expedition. This provides an excellent opportunity to examine the origin of the Kmin tilt. Comparison of the measured dips of the inclined bedding with the Kmin tilt of the AMS data shows that these two parameters are broadly consistent  from Unit I display erratic demagnetization trajectories. For the remaining 60 specimens, AF demagnetization, typically from ~20 to 40 mT, or thermal demagnetization, typically from 280 °C to 500 °C, effectively isolated the characteristic remanent magnetization (ChRM)  of the samples ranges from 3.10 × 10RM) Fig. . SamplesRM) Fig. . Thermal41 and the site was located at a low latitude region42, ChRMs with inclinations steeper than 70° may be affected by drilling-induced remagnetization41 and are rejected. Third, since the cores were drilled with the rotary core barrel (RCB) and were not fully oriented, we only use downhole changes in inclination to construct the magnetostratigraphy. The ChRMs data are summarized in Table We use the following criteria to select the ChRMs. First, ChRMs with a maximum angular deviation (MAD) greater than 16° are deemed unreliable and are excluded from further analysis. Second, given that drilling can induce a near-vertical overprintmin tilt approximates the steepness of tilted strata, tilt correction was performed by restoring Kmin axes to the vertical. An inclination-only fold test shows that ks/kg = 1.33, where ks and kg are the precision parameters in stratigraphic and geographic coordinates respectively, suggesting that the grouping of inclinations is improved after tilt correction. Therefore, the ChRM is likely primary in origin and the tilt-corrected inclination can be used to establish magnetostratigraphy. The assignment of magnetozones was aided by the biostratigraphic results from this core. Biostratigraphic data show that the nannofossil Zone NN18 is well defined with its upper boundary at ~505 mbsf and the lower boundary at ~790 mbsf, respectively8. The upper and lower boundary of Zone NN18 corresponds to 1.93 Ma and 2.39 Ma, respectively8. With this constraint, the predominant positive inclination values between ~470 mbsf and ~560 mbsf (N1) are assigned to a normal polarity zone corresponding to the Subchron C2n (Olduvai), which ranges from 1.778 to 1.945 Ma43  and 1.95 Ma (~720 mbsf) in the upper part of the relatively strong deformation zone7  and the upper slope (Site U1379) of the forearc Fig. . At uppene7 Fig. . Assumin379 Fig.  supports8, indicating a zone of relatively strong deformation. Also, the foliations are largely shallower than the bedding within this interval8, suggesting strong sub-horizontal compressional deformation at the frontal prism site. The upper bound age of the strong deformation zone is Pleistocene, around 1.89 Ma based on the biostratigraphic data8, which is similar to the estimated upper bound ages of the strong deformation zones at Sites U1380 (~1.83 Ma) and U1379 (~1.91 Ma), respectively.At frontal prism Site U1412, abundant faults, fractures, and foliations occur in Unit III at Site U1412 (~330 to 370 mbsf)8 in the relatively strong deformation zone at Site U1379 may indicate relatively moderate deformation in the upper slope. On the other hand, lithological difference may also contribute to the structural difference across the forearc. Nevertheless, the deformation of the stratigraphically coeval sediments is significant. The shell fragment-bearing sandstones underlying the relatively strong deformation zones mark shelf facies deposited in the uplifted forearc upon the arrival of the CR at the MAT7. The overlying deep-water facies that occupies the deformation zones represents deposition during the subsequent rapid forearc subsidence7 driven by the sudden, effective, and dramatic subduction erosion of the CR. In this context, the strong brief deformation of the deep-water facies at ~1.80 Ma across the forearc is best explained by the intense sub-horizontal shear immediately following the rapid forearc subsidence. Therefore, the widespread, brief, and relatively strong deformation is likely an integral part of the consequences in response to the sudden, dramatic subduction erosion associated with the near-flat subduction of the CR at the Costa Rican margin. Since subduction erosion causes the landward migration of the trench and subsidence in the forearc area in an erosive convergent margin, similar styles of deformation are probably common in other erosive convergent margins as well. The findings thus contribute to improved understanding of the crustal material transfer, hydrothermal fluid migration, and seismogenesis in erosive convergent margins.Collectively, these data indicate an interval of sediments spanning ~2.0 to ~1.83 Ma across the forearc that likely underwent a wholesale brief deformation at ~1.80 Ma. Also, the intensity of deformation appears to decrease from the trench to the upper slope according to the depicted deformation model Fig. . The occA total of 353 standard cylindrical paleomagnetic specimens were drilled from the working halves of sediment cores retrieved from IODP Expedition 344 Hole U1380C. Among these specimens, 60 were selected for shipboard studies, which included stepwise alternating field (AF) or thermal demagnetization up to 120 mT and 475 °C, respectively, to isolate the characteristic remanent magnetization (ChRM). The remaining specimens were investigated on shore. Most specimens from the upper part of the hole disintegrated during shipment, only 193 specimens arrived intact. Their low-field magnetic susceptibility (MS) and anisotropy of magnetic susceptibility (AMS) were first measured with an AGICO Kappabridge KLY-3. About half of these specimens were subsequently subjected to stepwise AF demagnetization up to 50 to 100 mT at increments of 5 or 10 mT using a Molspin demagnetizer. The other half of the specimens were thermally demagnetized using an ASC TD-48 thermal demagnetizer at increments of 30° to 50 °C, typically in 20 steps, up to 600 °C. Magnetic remanence was measured with a 2G Enterprises Inc. 755 superconducting rock magnetometer housed in a magnetically shielded room  in the Paleomagnetism Laboratory of Nanjing University, China.44 and were analyzed using principal component analysis45. The software PuffinPlot46 and PaleoMac47 were used to analyze the data and produce figures.The demagnetization data were presented graphically using vector endpoint plotsThe AMS, bedding dip, and ChRM data are summarized in Tables The data are provided in the Supplementary Information.Supplementary Information