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Partial purification and characterization of an acid dipeptide naphthylamidase (carboxytripeptidase) of the rat skin

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Zusammenfassung

Eine Peptidase mit der Fähigkeit Dipeptide aus Dipeptidnaphthylamiden freizusetzen, wurde aus Rattenhauthomogenat durch Präzipitation mit neutralen Salzen, Gelfiltration an Sephadex G-100 und CM- und DEAE-Cellulose-Chromatographie partiell getrennt. Die Peptidase hydrolysierte besonders Naphthylamide von Leucyl-Alanin und Lysyl-Alanin aber auch von Alanyl-Alanin und Glycyl-Prolin. Alanyl-Alanyl-Alanin wurde ins Alanyl-Alanin und freies Alanin hydrolysiert. Alanyl-Alanin und Alanyl-Alanyl-Alanyl-Alanin waren nicht gespalten. Die enzymatische Hydrolyse war optimal bei pH 5,3–5,7 und wurde durch SH-Gruppen oder Metallionen nicht aktiviert oder inhibiert. Das Enzym wurde bei 55° C inaktiviert.

Summary

A peptidase capable of liberating dipeptides from dipeptide naphthylamides, preferentially from naphthylamides of leucyl-alanine, lysyl-alanine, alanyl-alanine and glycyl-proline, was partially purified from rat skin homogenate. Purification procedure involved neutral salt precipitation, acid treatment, gel filtration on Sephadex G-100, and chromatographies on CM- and DEAE-cellulose. The enzyme was optimally active at pH 5.3–5.7 and was not effected by sulfhydryl reagents, metal ions nor chelators. High ionic concentration was found to be inhibitory. The enzyme was rapidly destroyed at 55° C. Alanyl-alanyl-alanine was found to be hydrolysed to alanyl-alanine and free alanine. Analyl-alanine and alanyl-alanyl-alanyl-alanine were not hydrolysed. The enzyme is suggested to correspond to the called dipeptidyl arylamidase II earlier described in bovine pituitary gland.

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Reference

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Authors and Affiliations

  1. Departments of Anatomy and Dermatology, University of Turku, Finlad

    Väinö K. Hopsu-Havu, Christer T. Jansén & Mikko Järvinen

Authors
  1. Väinö K. Hopsu-Havu
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  2. Christer T. Jansén
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  3. Mikko Järvinen
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Additional information

This report forms part V in our series on “Peptidases in the skin”, and is a part of the research project of the Skin Biology Research Unit in Turku (SBRU). Financial support was obtained as grants from the Sigrid Juselius Foundation and from the Finnish Medical Council to the senior author (V. K. H.-H.).

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Hopsu-Havu, V.K., Jansén, C.T. & Järvinen, M. Partial purification and characterization of an acid dipeptide naphthylamidase (carboxytripeptidase) of the rat skin. Arch. klin. exp. Derm. 236, 282–296 (1970). https://doi.org/10.1007/BF00508328

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  • DOI: https://doi.org/10.1007/BF00508328

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