Zolotarev Iu A, Sokolov O Iu, Kost N V, Vas'kovskiĭ B V, Miasoedov N F, Zozulia A A
Bioorg Khim. 2004 May-Jun;30(3):234-40. doi: 10.1023/b:rubi.0000030126.09208.c3.
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 approximately 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. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru.
开发了一种基于应用在所有氨基酸残基上均用氚普遍标记的亮氨酸脑啡肽来分析血浆中脑啡肽酶活性的方法。该方法能够同时对微量组织中的几种肽酶活性进行评估。[G-3H]亮氨酸脑啡肽通过固相催化同位素交换法制备（120 Ci/mmol），并通过血浆处理进行蛋白水解。所得放射性代谢产物在未标记的亮氨酸脑啡肽片段混合物作为内标的情况下通过高效液相色谱法进行分离。结果表明，氨肽酶、二肽基氨肽酶和二肽基羧肽酶分别对总酶活性的约80%、2%和10%负责。发现了羧肽酶降解亮氨酸脑啡肽的新途径，其占总脑啡肽降解活性的约6%。结果表明，贝司他汀主要抑制氨肽酶和羧肽酶，而赛兰克对羧肽酶和二羧肽酶更具特异性。该论文的英文版本：《俄罗斯生物有机化学杂志》，2004年，第30卷，第3期；另见http://www.maik.ru。
