Leu-Enkephalin Generally Labeled with Tritium in Studying the Selank Inhibiting Effect on the Enkephalin-Degrading Enzymes of Human Blood Plasma

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• 作者：Yu. A. Zolotarev ; O. Yu. Sokolov ; N. V. Kost ; B. V. Vaskovsky ; N. F. Myasoedov and A. A. Zozulya
• 关键词：bestatin ; biodegradation of peptides ; blood serum enkephalins ; enkephalins ; inhibitors of proteases ; selank
• 刊名：Russian Journal of Bioorganic Chemistry
• 出版年：2004
• 出版时间：2004
• 年：2004
• 卷：30
• 期：3
• 页码：207-212
• 全文大小：82 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Biochemistry
  Bioorganic Chemistry
  Organic Chemistry
  Biomedicine
  Russian Library of Science
  
• 出版者：MAIK Nauka/Interperiodica distributed exclusively by Springer Science+Business Media LLC.
• ISSN：1608-330X

文摘

A method of analysis of enkephalinase activity in blood plasma based on the application of Leu-enkephalin generally labeled with tritium at all its amino acid residues was developed. The method allows the simultaneous estimation of activity of several peptidases in microquantities of tissues. [G-3H]Leu-enkephalin was prepared by the method of solid phase catalytic isotope exchange (120 Ci/mmol) and subjected to proteolysis by the treatment with blood plasma. The resulting radioactive metabolites were separated by HPLC in the presence of the mixture of unlabeled fragments of Leu-enkephalin as internal standards. It was shown that aminopeptidases, dipeptidylaminopeptidases, and dipeptidylcarboxypeptidases respond for approximately 80%, 2%, and 10% of the total enzymatic activity, respectively. The new pathway of degradation of Leu-enkephalin by carboxypeptidase that provides for 6% of the total enkephalin-degrading activity was discovered. Bestatin was shown to predominantly inhibit aminopeptidases and carboxypeptidases, whereas selank is more specific for carboxypeptidases and dicarboxypeptidases.