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Archives of Dermatological Research

, Volume 255, Issue 3, pp 317–330 | Cite as

Human skin proteases

Separation and characterization of two acid proteases resembling cathepsin B1 and cathepsin D and of an inhibitor of cathepsin B1
  • Jorma E. Fräki
Article

Summary

Two acid proteases, one hydrolysing hemoglobin and the other hydrolysing benzoyl arginine naphthyamide (BANA), were separated and partially purified from human skin buffer extract.

The acid protease hydrolysing hemoglobin was purified about 190 fold by Sephadex G-100 gel filtration and DEAE-cellulose chromatography. It hydrolysed hemoglobin at pH 3.5, casein at pH 5.8 and skin protein substrate at pH 6.0. It did not markedly hydrolyse synthetic protease substrates. The molecular size of this protease was 38000. The protease was insensitive to common protease modifiers and closely resembles cathepsin D purified from other organs.

The BANA-hydrolysing acid protease was purified about 760 fold by Sephadex G-100 gel filtration and affinity chromatography on organomercurial Sepharose 4B gel. It preferentially hydrolysed BAEE, BANA and BAA with an optimum at pH 5.8. The hydrolysis of BAPA, LeuNA and protein substrates was very low. This acid protease was found to be highly dependent on reducing agents, as DTT, and chelating agents, as EDTA, and was inhibited by pCMB and TLCK. The molecular size of the enzyme was 28000. This protease closely resembles cathepsin B1 purified from other organs.

Human skin was also shown to contain a low activity of benzoyl arginine amide (BAA) hydrolysing acid protease with a molecular size of about 50000 and resembling cathepsin B2.

Human skin contained an inhibitor with a molecular size of about 13000 against human skin cathepsin B1. This inhibitor did not inhibit trypsin, chymotrypsin or skin proteases other than cathepsin B1.

Keywords

Human Skin Molecular Size Chymotrypsin Acid Protease Skin Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Zusammenfassung

Zwei saure Proteasen, von denen die eine Hämoglobin und die andere Benzoylarginin-naphthylamid (BANA) spalten, wurden aus menschlichem Hautextrakt abgetrennt und teilweise gereinigt.

Die saure Protease, die Hämoglobin spaltet, wurde ca. 190mal mit Sephadex G-100 Gel Filtration und DEAE-Cellulose-Chromatographie gereinigt. Das Enzym hydrolyziert Hämoglobin bei einem pH-Optimum von 3,5, Casein bei demjenigen von 5,8 und Dermoproteinsubstrat bei einem pH-Optimum von 6,0. Die synthetischen Substrate wurden nicht hydrolyziert. Die Molekülgröße dieser Protease war 38000. Die Protease war unempfindlich gegen gewöhnliche Modifizierer und ähnelte stark Kathepsin D, das aus anderen Organen gereinigt worden ist.

Die sauere Protease, die BANA spaltet, wurde ca. 760mal mit Sephadex G-100 Gel Filtration und Affinitätschromatographie mit organomerkurischen Sepharose 4 B Gel gereinigt. Das Enzym hydrolyziert eher BAEE, BANA und BAA bei einem pH-Optimum von 5,8. BAPA und LeuNA wurden sehr langsam gespaltet. Diese sauere Protease war sehr abhängig von reduzierenden Agenten, wie DTT und von chelatierenden Agenten, wie EDTA, und wurde mit pCMB und TLCK gehemmt. Die Molekülgröße dieses Enzyms war 28000. Diese Protease ähnelte stark Kathepsin B1, das aus anderen Organen gereinigt worden ist.

In der menschlichen Haut wurde auch eine Protease nachgewiesen, die Benzoylarginin-amid (BAA) hydrolyziert, eine Molekülgröße ca. 50000 hat und ähnlich dem Kathepsin B2 ist.

Die menschliche Haut enthielt einen Inhibitor von der Molekülgröße ca. 13000 gegen das Kathepsin B1 aus menschlicher Haut. Inhibitor hemmte nicht Trypsin, Chymotrypsin oder die anderen Proteasen der Haut außer Kathepsin B1.

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Copyright information

© Springer-Verlag 1976

Authors and Affiliations

  • Jorma E. Fräki
    • 1
    • 2
  1. 1.Department of DermatologyUniversity of TurkuFinland
  2. 2.Institute of BiomedicineUniversity of TurkuFinland

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