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 Wojcik, Sherwin Wilk, Satoshi Omura, Leszek Kozlowski, Tomasz Stoklosa, Krzysztof Worowski, Piotr Radziwon

Research output: Contribution to journalArticle

40 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Cathepsin A
Proteasome Inhibitors
Lactones
Proteasome Endopeptidase Complex
Chymotrypsin
Enzymes
Platelets
Blood Platelets
Carboxypeptidases
lactacystin
Concanavalin A
Antigens

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Separation of cathepsin A-like enzyme and the proteasome : Evidence that lactacystin/β-lactone is not a specific inhibitor of the proteasome. / Ostrowska, Halina; Wojcik, Cezary; Wilk, Sherwin; Omura, Satoshi; Kozlowski, Leszek; Stoklosa, Tomasz; Worowski, Krzysztof; Radziwon, Piotr.

In: International Journal of Biochemistry and Cell Biology, Vol. 32, No. 7, 01.07.2000, p. 747-757.

Research output: Contribution to journalArticle

Ostrowska, Halina ; Wojcik, Cezary ; Wilk, Sherwin ; Omura, Satoshi ; Kozlowski, Leszek ; Stoklosa, Tomasz ; Worowski, Krzysztof ; Radziwon, Piotr. / Separation of cathepsin A-like enzyme and the proteasome : Evidence that lactacystin/β-lactone is not a specific inhibitor of the proteasome. In: International Journal of Biochemistry and Cell Biology. 2000 ; Vol. 32, No. 7. pp. 747-757.
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