Zolotarev Yu A, Dolotov O V, Inozemtseva L S, Dadayan A K, Dorokhova E M, Andreeva L A, Alfeeva L Yu, Grivennikov I A, Myasoedov N F
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia.
Amino Acids. 2006 Jun;30(4):403-8. doi: 10.1007/s00726-006-0328-8. Epub 2006 May 26.
Here a new approach of the elucidation of paths of proteolytic biodegradation of physiologically active peptides, based on the use of a peptide with isotopic label at all amino acid residues and the enrichment of HPLC samples with unlabeled peptide fragments in UV-detectable concentration, has been proposed. The method has been applied for the investigation of degradation dynamics of the neuroactive heptapeptide MEHFPGP (Semax) in the presence of plasma membranes, and cultures of glial and neuronal cells obtained from the rat basal forebrain. The splitting away of ME and GP, and formation of pentapeptides are the predominant processes in the presence of all tested objects, whereas the difference in patterns of resulting peptide products for glial and neuronal cells has been detected. In conclusion, the approach applied allows analyzing physiologically active peptide concentrations in biological tissues and degradation pathways of peptides in the presence of targets of their action.
本文提出了一种阐明生理活性肽蛋白水解生物降解途径的新方法，该方法基于使用所有氨基酸残基均带有同位素标记的肽，并通过未标记的肽片段以可紫外检测的浓度富集高效液相色谱（HPLC）样品。该方法已应用于研究神经活性七肽MEHFPGP（Semax）在质膜以及从大鼠基底前脑获得的神经胶质细胞和神经元细胞培养物存在下的降解动力学。在所有测试对象存在的情况下，ME和GP的裂解以及五肽的形成是主要过程，而神经胶质细胞和神经元细胞产生的肽产物模式存在差异。总之，所应用的方法能够分析生物组织中生理活性肽的浓度以及在其作用靶点存在的情况下肽的降解途径。
