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PMID: 8836139 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Binding of ATP and ATP analogues to the uncoating ATPase Hsc70 (70 kDa heat-shock cognate protein).

The Biochemical journal ·Vol. 318 ( Pt 3) ·1996-09-15 ·Pages 923-9

Buxbaum E, Woodman PG

Abstract

Nucleotide binding to the 70 kDa heat-shock cognate protein (Hsc70) from mung bean seeds and pig brain was investigated, as well as the clathrin uncoating activity of Hsc70 in the presence of these nucleotides. The two enzymes were found to behave identically. ATP bound to two different forms of Hsc70, with dissociation constants of 1.1 +/- 0.1 microM and 1.4 +/- 0.7 mM respectively at 25 degrees C. This corresponds to delta G0' = -34 and -16 kJ/mol respectively. From the temperature-dependence of the dissociation constant of the high-affinity site, delta H0' was calculated to -36 +/- 2 kJ/mol. This gives delta S0' = 6.7 J/mol per K. Adenosine 5'-[gamma-thio]triphosphate, ADP, adenosine 5'-[beta, gamma-imino]triphosphate and adenosine 5'-[beta, gamma-methylene]triphosphate showed dissociation constants of 2.3, 11, 31 and 284 microM respectively. The order of affinities corresponded to the order of effectiveness in uncoating of pig brain coated vesicles. The implications of these findings for the mechanism of Hsc70 action are discussed.

MeSH Terms
Adenosine Triphosphatases/chemistry,metabolism Adenosine Triphosphate/analogs & derivatives,metabolism Animals Binding Sites Brain/metabolism Carrier Proteins/chemistry,metabolism Clathrin/metabolism Coated Pits, Cell-Membrane/metabolism Fabaceae/metabolism HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins In Vitro Techniques Kinetics Plants, Medicinal Swine Thermodynamics
Chemicals
Carrier Proteins Clathrin HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins Adenosine Triphosphate Adenosine Triphosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Buxbaum E
School of Biological Science, University of Manchester, U.K.
Woodman P G
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31 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3180923
Published
1996-09-15
Pages
923-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1217706
Subset
IM
External Links
PubMed source DOI: 10.1042/bj3180923 PMC Full Text View journal details
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