Separation of cathepsin A-like enzyme and the proteasome: Evidence that lactacystin/β-lactone is not a specific inhibitor of the proteasome

Halina Ostrowska, Cezary Wójcik, Sherwin Wilk, Satoshi Omura, Leszek Kozlowski, Tomasz Stoklosa, Krzysztof Worowski, Piotr Radziwon

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Previous studies have described a human platelet cathepsin A-like enzyme with a number of similarities to the 'acidic' and 'neutral' chymotrypsin-like activities of the proteasome. This includes its strong inhibition by the highly specific proteasome inhibitor Lactacystin/β-lactone, suggesting that either the Cbz-Phe-Ala-hydrolyzing activity attributed to cathepsin A was due to the chymotrypsin-like activity of the proteasome or that lactacystin was not a specific inhibitor of the proteasome. In the present study we discard the first possibility on the basis of the following findings: (a) human platelet cathepsin A, unlike proteasome, binds to concanavalin A, and does not bind to Heparin-Sepharose at pH 7.4; (b) neither the chymotrypsin-like activity of the proteasome, nor proteasome antigens are detected in the cathepsin A preparation; (c) purified proteasome does not exhibit Cbz-Phe- Ala-hydrolyzing activity; (d) Z-Ile-Glu-(Ot-Bu)Ala-leucinal (PSI), a compound that selectively inhibits the chymotrypsin-like activity of the proteasome at a concentration of 10 μM has no inhibitory effect on the carboxypeptidase activity of cathepsin A; (e) cathepsin A, free of the proteasome, is completely inhibited by micromolar concentrations of lactacystin/β-lactone. It is therefore concluded that lactacystin/β-lactone is not a specific inhibitor of the proteasome. (C) 2000 Elsevier Science Ltd.

Original languageEnglish (US)
Pages (from-to)747-757
Number of pages11
JournalInternational Journal of Biochemistry and Cell Biology
Volume32
Issue number7
DOIs
StatePublished - Jul 1 2000

Keywords

  • Cathepsin A
  • Human platelets
  • Lactacystin
  • PSI
  • Proteasome

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

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