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PMID: 2557825 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Ubiquitin-protein conjugates accumulate in the lysosomal system of fibroblasts treated with cysteine proteinase inhibitors.

The Biochemical journal ·Vol. 263 ·No. 1 ·1989-10-01 ·Pages 47-55

Doherty FJ, Osborn NU, Wassell JA, Heggie PE, Laszlo L, Mayer RJ

Abstract

Mouse fibroblasts (3T3-L1 cells) accumulate detergent- and salt-insoluble aggregates of proteins conjugated to ubiquitin when incubated in the presence of inhibitors of lysosomal cysteine cathepsins, including E-64. These ubiquitin-protein conjugates co-fractionate with lysosomes on density gradients and are found in multivesicular dense bodies which by electron microscopy appear to be engaged in microautophagy. Both E-64 and ammonium chloride increase the intracellular concentration of free ubiquitin, but only E-64 leads to the formation of insoluble lysosomal ubiquitin-protein conjugates. The results are discussed in relation to the possible intracellular roles of ubiquitin conjugation.

MeSH Terms
Acid Phosphatase/analysis Animals Blotting, Western Cells, Cultured Cysteine Proteinase Inhibitors/pharmacology Electrophoresis, Polyacrylamide Gel Fibroblasts/drug effects,enzymology Hydrolysis Immunohistochemistry L-Lactate Dehydrogenase/analysis Lysosomes/metabolism Mice Microscopy, Electron Proteins/metabolism Ubiquitins/metabolism
Chemicals
Cysteine Proteinase Inhibitors Proteins Ubiquitins L-Lactate Dehydrogenase Acid Phosphatase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Doherty F J
Department of Biochemistry, University of Nottingham Medical School, Queen's Medical Centre, U.K.
Osborn N U
Wassell J A
Heggie P E
Laszlo L
Mayer R J
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2630047
Published
1989-10-01
Pages
47-55
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1133389
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2630047 PMC Full Text View journal details
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