Ligand fishing of anti neurodegenerative components from Lonicera japonica using magnetic nanoparticles immobilised with monoamine oxidase B.
In this work, monoamine oxidase B was immobilised onto magnetic nanoparticles to prepare a new type of affinity solid phase extraction adsorbent, which was used to extract the possible anti neurodegenerative components from the Lonicera japonica flower extracts. Coupled with high performance liquid chromatography with mass spectrometry, two monoamine oxidase B ligands were fished out and identified as isochlorogenic acid A and isochlorogenic acid C, which were found to be inhibitors of the enzyme for the first time, with similar half maximal inhibitory concentration values of 29.05 0.49 and 29.77 1.03 mM, respectively. Furthermore, equilibrium dialysis dissociation assay of enzyme inhibitor complex showed that both compounds have reversible binding patterns to monoamine oxidase B, and kinetic analysis demonstrated that they were mixed type inhibitors for monoamine oxidase B, with Ki and Kis values of 9.55 and 37.24 mM for isochlorogenic acid A, 9.53 and 35.50 mM for isochlorogenic acid C, respectively. The results indicated that isochlorogenic acid A and isochlorogenic acid C were the major active components responsible for the anti degenerative activity of the flowers of L. japonica, while magnetic nanoparticles immobilised monoamine oxidase B could serve as an efficient solid phase extraction adsorbent to specifically extract monoamine oxidase B inhibitors from complex herbal extracts.
Journal of separation science