A specific tripeptidyl substrate for tripeptidyl peptidase activity is effectively hydrolyzed by alanyl aminopeptidase/aminopeptidase N/CD13 in the rat kidney

Q4 Medicine

Archives of histology and cytology Pub Date : 2016-03-03 DOI:10.1679/AOHC.76.1

M. Shibata, M. Koike, S. Kusumi, Noboru Sato, Y. Uchiyama

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引用次数: 1

Abstract

amino terminal amino acid sequence of the peptidase was x-Ala-Pro-x-Leu-Pro-Gly-Ser-Thr-Ser-Ala-Thr-x-x-Ser, where x indicates undetectable amino acid residues, and the antiserum against the peptidase was immunopositive for the brush border of a renal proximal tubule and the small intestine, and the surface membrane of bile canaliculi. These results indicate that the unknown peptidase that hydrolyzed AAF-MCA is the soluble form of aminopeptidase N/CD13, and caution is required when using AAF-MCA as a substrate for tripeptidyl peptidase assays. Summary. L-Alanyl-L-alanyl-L-phenylalanine 4-methyl-coumaryl-7-amide (AAF-MCA) is one of the classic substrates for use with tripeptidyl peptidases (TPP-I and TPP-II). We have previously clarified the tissue distribution of TPP-I in detail and noted that the protein expression of TPP-I is often incompatible with its enzyme activity. Herein, we describe the unknown peptidase, which could effectively hydrolyze AAF-MCA, in the rat kidney. The peptidase was purified after four chromatography steps, and its enzyme characteristics were elucidated. The peptidase activity was inhibited by amastatin, bestatin, and o-phenanthroline and was also inhibited by zinc and copper ions. The substrate specificity for several monoamino acidic-MCAs revealed that the peptidase had an affinity for alanyl-MCA. The

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在大鼠肾脏中，丙氨酰氨基肽酶/氨基肽酶N/CD13可有效水解三肽酶活性的特异性三肽基底物

肽酶氨基末端氨基酸序列为x- ala - pro -x- leu - pro - gly - ser - thr - ser - ala - thr -x-x- ser，其中x表示未检测到氨基酸残基，抗血清对肾近端小管、小肠刷状缘和胆管表面膜均呈免疫阳性。这些结果表明，水解AAF-MCA的未知肽酶是氨基肽酶N/CD13的可溶性形式，使用AAF-MCA作为三肽基肽酶测定的底物时需要谨慎。总结。l-丙烯酰- l-丙烯酰- l-苯丙氨酸4-甲基-香豆醇-7-酰胺(AAF-MCA)是三肽基肽酶(TPP-I和TPP-II)的经典底物之一。我们之前已经详细阐明了tpp - 1的组织分布，并指出tpp - 1的蛋白表达往往与其酶活性不相容。在此，我们描述了一种未知的肽酶，它可以有效地水解大鼠肾脏中的AAF-MCA。经四层析纯化该肽酶，并对其酶学特性进行了分析。肽酶活性受阿马伐他汀、百司他汀和邻菲罗啉的抑制，锌离子和铜离子也有抑制作用。对几种单氨基酸- mca的底物特异性表明该肽酶对丙烯酰- mca具有亲和力。的

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来源期刊

Archives of histology and cytology 生物-细胞生物学

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期刊介绍： The Archives of Histology and Cytology provides prompt publication in English of original works on the histology and histochemistry of man and animals. The articles published are in principle restricted to studies on vertebrates, but investigations using invertebrates may be accepted when the intention and results present issues of common interest to vertebrate researchers. Pathological studies may also be accepted, if the observations and interpretations are deemed to contribute toward increasing knowledge of the normal features of the cells or tissues concerned. This journal will also publish reviews offering evaluations and critical interpretations of recent studies and theories.