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PMID: 20974815 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Quality control for unfolded proteins at the plasma membrane.

The Journal of cell biology ·Vol. 191 ·No. 3 ·2010-11-01 ·Pages 553-70

Apaja PM, Xu H, Lukacs GL

Abstract

Cellular protein homeostasis profoundly depends on the disposal of terminally damaged polypeptides. To demonstrate the operation and elucidate the molecular basis of quality control of conformationally impaired plasma membrane (PM) proteins, we constructed CD4 chimeras containing the wild type or a temperature-sensitive bacteriophage λ domain in their cytoplasmic region. Using proteomic, biochemical, and genetic approaches, we showed that thermal unfolding of the λ domain at the PM provoked the recruitment of Hsp40/Hsc70/Hsp90 chaperones and the E2-E3 complex. Mixed-chain polyubiquitination, monitored by bioluminescence resonance energy transfer and immunoblotting, is responsible for the nonnative chimera-accelerated internalization, impaired recycling, and endosomal sorting complex required for transport-dependent lysosomal degradation. A similar paradigm prevails for mutant dopamine D4.4 and vasopressin V2 receptor removal from the PM. These results outline a peripheral proteostatic mechanism in higher eukaryotes and its potential contribution to the pathogenesis of a subset of conformational diseases.

MeSH Terms
Animals Bacteriophage lambda/chemistry CD4 Antigens/chemistry,metabolism COS Cells Cell Line Cell Membrane/chemistry,metabolism Chlorocebus aethiops Chromatin Immunoprecipitation Endocytosis Endosomal Sorting Complexes Required for Transport/metabolism Green Fluorescent Proteins/chemistry,metabolism Humans Lysosomes/metabolism Molecular Chaperones/metabolism Multivesicular Bodies/metabolism Protein Conformation Protein Denaturation Protein Unfolding Receptors, G-Protein-Coupled/chemistry,metabolism Temperature Ubiquitination
Chemicals
CD4 Antigens Endosomal Sorting Complexes Required for Transport Molecular Chaperones Receptors, G-Protein-Coupled enhanced green fluorescent protein Green Fluorescent Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Apaja Pirjo M
Department of Physiology, McGill University, Montréal, Quebec, Canada.
Xu Haijin
Lukacs Gergely L
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201006012
Published
2010-11-01
Epub
2010-00-25
Pages
553-70
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3003321
Subset
IM
Grants
Canadian Institutes of Health Research · Canada
External Links
PubMed source DOI: 10.1083/jcb.201006012 PMC Full Text View journal details
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