Signal and binding. II. Converting physico-chemical responses to macromolecule-ligand interactions into thermodynamic binding isotherms.
Physico-chemical titration techniques are the most commonly used methods in characterizing molecular interactions. These methods are mainly based on spectroscopic, calorimetric, hydrodynamic, etc., measurements. However, truly quantitative physico-chemical methods are absolutely based on the determination of the relationship between the measured signal and the total average degree of binding in order to obtain meaningful interaction parameters. The relationship between the observed physico-chemical signal of whatever nature and the degree of binding must be determined and not assumed, based on some ad hoc intuitive relationship/model, leading to determination of the true binding isotherm. The quantitative methods reviewed and discussed here allow an experimenter to rigorously determine the degree of binding and the free ligand concentration, i.e., they lead to the construction of the thermodynamic binding isotherm in a model-independent fashion from physico-chemical titration curves.
Thermodynamic extent of counterion release upon binding oligolysines to single-stranded nucleic acids.
A major contribution to the binding free energy associated with most protein-nucleic acid complexes is the increase in entropy due to counterion release from the nucleic acid that results from electrostatic interactions. To examine this quantitatively, we have measured the thermodynamic extent of counterion release that results from the interaction between single-stranded homopolynucleotides and a series of oligolysines, possessing net charges z = 2-6, 8, and 10. This was accomplished by measuring the salt dependence of the intrinsic equilibrium binding constants--i.e., (delta log Kobs/delta log[K+])--over the range from 6 mM to 0.5 M potassium acetate. These data provide a rigorous test of linear polyelectrolyte theories that have been used to interpret the effects of changes in bulk salt concentration on protein-DNA binding equilibria, since single-stranded nucleic acids have a lower axial charge density than duplex DNA. Upon binding to poly(U), the thermodynamic extent of counterion release per oligolysine charge, z, is 0.71 +/- 0.03, which is significantly less than unity and less than that measured upon binding duplex DNA. These results are most simply interpreted using the limiting law predictions of counterion condensation and cylindrical Poisson-Boltzmann theories, even at the high salt concentrations used in our experiments. Accurate estimates of the thermodynamic extent of counterion binding and release for model systems such as these facilitate our understanding of the energetics of protein-nucleic acid interactions. These data indicate that for simple oligovalent cations, the number of ionic interactions formed in a complex with a linear nucleic acid can be accurately estimated from a measure of the salt dependence of the equilibrium binding constant, if the thermodynamic extent of ion release is known.
Students as promoters of rural extension in aquaculture in the Marajó archipelago, Pará, Brazil
Aquaculture is the creation of aquatic organisms. An expanding activity, being of great importance its approach in rural extension. Thus, the objective of this study was to analyze the contribution of extension activities in aquaculture in the professional training of students in Marajó, Amazon, Brazil. The project “Technology transfer through training and technical assistance for tambaqui breeders, Colossoma macropomum, in the archipelago of Marajó/PA” was developed by 10 technician and under graduate students from rural education course and the technical course in aquaculture from the Federal Institute of Education, Science and Technology of Pará (IFPA) Campus Breves, in the city of Curralinho. They participated in the stages of data collection in aquaculture enterprises, training for producers and monitoring of aquaculture production, acting as researchers, instructors and extension workers, respectively. The duration of the project was from August 2019 to March 2020. At the end of the project, a questionnaire was applied to the students involved. The results showed that the aquaculture extension activities developed were relevant and significant, for the construction of knowledge in a mutual way between those involved (students and aquaculture producers). The acquired experiences will substantiate and subsidize the professional practice of these students in a concrete and effective way. In conclusion, the extension project represents an excellent tool in the teaching-learning process for students.