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PMID: 3020549 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Endogenous inhibitor of nonlysosomal high molecular weight protease and calcium-dependent protease.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 20 ·1986-10-00 ·Pages 7588-92

Murakami K, Etlinger JD

Abstract

An endogenous inhibitor of high molecular weight protease was purified from human erythrocytes and partially characterized. The inhibitor was isolated by DEAE-Sephacel ion-exchange chromatography followed by separation on a Bio-Gel A-0.5m column. The inhibitor displayed a native Mr of 240,000 and contained a single subunit of Mr 40,000 after NaDodSO4/polyacrylamide gel electrophoresis. The Mr 240,000 hexamer inhibited high molecular weight protease noncompetitively (Ki = 8.3 X 10(-8) M) and showed marked susceptibility to proteolytic digestion and heat treatment. The purified factor was also a potent inhibitor of calcium-dependent protease (Ki = 2.8 X 10(-8) M), whereas it had no effect on trypsin, chymotrypsin, or papain. Heat treatment (50-70 degrees C X 10 min) caused loss of inhibition against high molecular weight protease; however, inhibition of calcium-dependent protease was stable under the same conditions. This result is consistent with different domains on the inhibitor that interact with high molecular weight protease and calcium-dependent protease. Together with earlier studies in which repression of inhibitor by an ATP-ubiquitin-dependent process was proposed, the present results suggest a general mechanism for regulation of multiple nonlysosomal proteases that are complexed with endogenous inhibitors.

MeSH Terms
Adenosine Triphosphate/pharmacology Animals Calpain/antagonists & inhibitors Humans Molecular Weight Protease Inhibitors/analysis,isolation & purification,pharmacology Rabbits
Chemicals
Protease Inhibitors Adenosine Triphosphate Calpain
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Murakami K
Etlinger J D
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44 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.20.7588
Published
1986-10-00
Pages
7588-92
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386766
Subset
IM
Grants
NHLBI NIH HHS · 1R01 HL 31494 · United States
NIADDK NIH HHS · IT 32 AM 07289 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.20.7588 PMC Full Text View journal details
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