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PMID: 8670798 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The human cytosolic molecular chaperones hsp90, hsp70 (hsc70) and hdj-1 have distinct roles in recognition of a non-native protein and protein refolding.

The EMBO journal ·Vol. 15 ·No. 12 ·1996-06-17 ·Pages 2969-79

Freeman BC, Morimoto RI

Abstract

The properties of molecular chaperones in protein-assisted refolding were examined in vitro using recombinant human cytosolic chaperones hsp90, hsc70, hsp70 and hdj-1, and unfolded beta-galactosidase as the substrate. In the presence of hsp70 (hsc70), hdj-1 and either ATP or ADP, denatured beta-galactosidase refolds and forms enzymatically active tetramers. Interactions between hsp90 and non-native beta-galactosidase neither lead to refolding nor stimulate hsp70- and hdj-1-dependent refolding. However, hsp90 in the absence of nucleotide can maintain the non-native substrate in a 'folding-competent' state which, upon addition of hsp70, hdj-1 and nucleotide, leads to refolding. The refolding activity of hsp70 and hdj-1 is effective across a broad range of temperatures from 22 degrees C to 41 degrees C, yet at extremely low (4 degrees C) or high (>41 degrees C) temperatures refolding activity is reversibly inhibited. These results reveal two distinct features of chaperone activity in which a non-native substrate can be either maintained in a stable folding-competent state or refolded directly to the native state; first, that the refolding activity itself is temperature sensitive and second, that hsp90, hsp70 (hsc70) and hdj-1 each have distinct roles in these processes.

MeSH Terms
Adenine Nucleotides/physiology Animals Baculoviridae/genetics Cell Line HSP40 Heat-Shock Proteins HSP70 Heat-Shock Proteins/metabolism,physiology HSP90 Heat-Shock Proteins/genetics,metabolism,physiology Heat-Shock Proteins Humans Molecular Chaperones/metabolism,physiology Protein Binding Protein Folding Recombinant Proteins/genetics,metabolism Spodoptera Temperature beta-Galactosidase/metabolism
Chemicals
Adenine Nucleotides DNAJB1 protein, human HSP40 Heat-Shock Proteins HSP70 Heat-Shock Proteins HSP90 Heat-Shock Proteins Heat-Shock Proteins Molecular Chaperones Recombinant Proteins beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Freeman B C
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL 60208, USA.
Morimoto R I
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1996-06-17
Pages
2969-79
Language
English
Region
England
NLM ID
8208664
PMCID
PMC450238
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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