The 90-kDa heat shock protein (hsp-90) possesses an ATP binding site and autophosphorylating activity

P. Csermely, C. Ronald Kahn

Research output: Contribution to journalArticle

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Abstract

The 90-kDa heat shock protein (hsp-90) is an abundant cytosolic protein believed to play a role in maintenance of protein trafficking and closely associated with several steroid hormone receptors. Incubation of highly purified hsp-90 with [γ-32P]ATP results in its autophosphorylation on serine residues. There are several lines of evidence which suggest that this activity is due to a kinase intrinsic to hsp-90 rather than some closely associated protein kinases: 1) the phosphorylation persists after the removal of casein kinase II by heparin chromatography and after immunoprecipitation of hsp-90 with anti-hsp-90 antibodies. 2) The approximate kM for ATP of the reaction is 0.16 mM, higher than that of many other protein kinases. 3) Phosphorylation is not affected by a number of activators and inhibitors of other known kinases which might associate with hsp-90. 4) The phosphorylation displays a unique cation dependence being most active in the presence of Ca2+ and practically inactive with Mg2+, although the autophosphorylation in the presence of Mg2+ is activated by histones and polyamines. 5) The activity is remarkably heat-stable; incubation of hsp-90 for 20 min at 95°C results in only a 60% decrease in autophosphorylation. 6) Finally, and most importantly, purified hsp-90 can be labeled with azido-ATP and it is able to bind to ATP-agarose. The binding shows similar cation dependence to the autophosphorylation. These data are in agreement with the presence of a consensus sequence for ATP binding sites in the primary structure of the protein similar to that observed in the 70-kDa heat-shock proteins. Our data suggest the 90-kDa heat shock protein possesses an enzymatic activity analogous in many respects to the similar activity of the 70-kDa heat shock proteins. This may represent an important, previously unrecognized function of hsp-90.

Original languageEnglish
Pages (from-to)4943-4950
Number of pages8
JournalJournal of Biological Chemistry
Volume266
Issue number8
Publication statusPublished - Mar 15 1991

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HSP90 Heat-Shock Proteins
Heat-Shock Proteins
Phosphorylation
Adenosine Triphosphate
Binding Sites
HSP70 Heat-Shock Proteins
Protein Kinases
Cations
Phosphotransferases
Steroid hormones
Casein Kinase II
Proteins
Steroid Receptors
Consensus Sequence
Polyamines
Protein Transport
Chromatography
Immunoprecipitation
Histones
Serine

ASJC Scopus subject areas

  • Biochemistry

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The 90-kDa heat shock protein (hsp-90) possesses an ATP binding site and autophosphorylating activity. / Csermely, P.; Kahn, C. Ronald.

In: Journal of Biological Chemistry, Vol. 266, No. 8, 15.03.1991, p. 4943-4950.

Research output: Contribution to journalArticle

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