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

• Shibata Masahiro

  Department of Morphological Sciences, Kagoshima University Graduate School of Medical and Dental Sciences,

• Koike Masato

  Department ofCell Biology and Neuroscience, Juntendo University Graduate School of Medicine,

• Kusumi Satoshi

  Department of Morphological Sciences, Kagoshima University Graduate School of Medical and Dental Sciences,

• Sato Noboru

  Division of Gross Anatomy and Morphogenesis,Department of Regenerative and Transplant Medicine,Niigata University Graduate School of Medical and Dental Sciences,

• Uchiyama Yasuo

  Department of Cellular and Molecular Neuropathology, Juntendo University Graduate School of Medicine,

Abstract

L-Alanyl-L-alanyl-L-phenylalanine 4-methylcoumaryl-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 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.

Journal

• Archives of Histology and Cytology

  Archives of Histology and Cytology 76 (1), 1-8, 2016

  International Society of Histology and Cytology

Keywords

• Histology

Details

• CRID: 1390001206413310080
• NII Article ID: 130005131324
• DOI: 10.1679/aohc.76.1
• ISSN: 13491717
• ISSN: 09149465
• Web Site: https://www.jstage.jst.go.jp/article/aohc/76/1/76_1/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2017381399
• Text Lang: en
• Data Source:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed