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PMID: 2820375 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Skeletal muscle and liver contain a soluble ATP + ubiquitin-dependent proteolytic system.

The Biochemical journal ·Vol. 243 ·No. 2 ·1987-04-15 ·Pages 335-43

Fagan JM, Waxman L, Goldberg AL

Abstract

Although protein breakdown in most cells seems to require metabolic energy, it has only been possible to establish a soluble ATP-dependent proteolytic system in extracts of reticulocytes and erythroleukemia cells. We have now succeeded in demonstrating in soluble extracts and more purified preparations from rabbit skeletal muscle a 12-fold stimulation by ATP of breakdown of endogenous proteins and a 6-fold stimulation of 125I-lysozyme degradation. However, it has still not been possible to demonstrate such large effects of ATP in similar preparations from liver. Nevertheless, after fractionation by DEAE-chromatography and gel filtration, we found that extracts from liver as well as muscle contain both the enzymes which conjugate ubiquitin to 125I-lysozyme and an enzyme which specifically degrades the ubiquitin-protein conjugates. When this proteolytic activity was recombined with the conjugating enzymes, ATP + ubiquitin-dependent degradation of many proteins was observed. This proteinase is unusually large, approx. 1500 kDa, requires ATP hydrolysis for activity and resembles the ubiquitin-protein-conjugate degrading activity isolated from reticulocytes. Thus the ATP + ubiquitin-dependent pathway is likely to be present in all mammalian cells, although certain tissues may contain inhibitory factors.

MeSH Terms
Adenosine Triphosphate/pharmacology Animals Chromatography, Gel Electrophoresis, Polyacrylamide Gel In Vitro Techniques Liver/drug effects,enzymology,metabolism Muscle Proteins/metabolism Muscles/drug effects,enzymology,metabolism Peptide Hydrolases/isolation & purification,metabolism Protease Inhibitors/pharmacology Proteins/metabolism Rabbits Ubiquitins/pharmacology
Chemicals
Muscle Proteins Protease Inhibitors Proteins Ubiquitins Adenosine Triphosphate Peptide Hydrolases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fagan J M
Waxman L
Goldberg A L
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65 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2430335
Published
1987-04-15
Pages
335-43
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1147859
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2430335 PMC Full Text View journal details
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