Preformed GroES oligomers are not required as functional cochaperonins

Jeffrey W. Seale, John Chirgwin, Borries Demeler, Paul M. Horowitz

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have previously shown that the C-terminal sequence of GroES is required for oligomerization [Seale and Horowitz (1995), J. Biol. Chem. 270, 30268-30270]. In this report, we have generated a C-terminal deletion mutant of GroES with a significantly destabilized oligomer and have investigated its function in the chaperonin- assisted protein folding cycle. Removal of the two C-terminal residues of GroES results in a cochaperonin [GroESD(96-97)] that is monomeric at concentrations where GroES function is assessed. Using equilibrium ultracentrifugation, we measured the dissociation constant for the oligomer-monomer equilibrium to be 7.3 x 10-34 M6. The GroESD(96-97) is fully active as a cochaperonin. This mutant is able to inhibit the ATPase activity of GroEL to levels comparable to wild-type GroES. It is also able to assist the refolding of urea-denatured rhodanese by GroEL. While GroESD(96- 97) can function at levels comparable to wild-type GroES, higher concentrations of mutant are required to produce the same effect. These results support the idea that the preformed GroES heptamer is not required for function, but they suggest that the oligomeric cochaperonin is most efficient.

Original languageEnglish (US)
Pages (from-to)661-668
Number of pages8
JournalJournal of Protein Chemistry
Volume16
Issue number7
DOIs
StatePublished - Oct 1997
Externally publishedYes

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Thiosulfate Sulfurtransferase
Chaperonins
Ultracentrifugation
Protein Folding
Oligomers
Adenosine Triphosphatases
Urea
Protein folding
Oligomerization
Monomers

Keywords

  • ATPase
  • Chaperonins
  • GroEL
  • GroES
  • Protein folding

ASJC Scopus subject areas

  • Biochemistry

Cite this

Preformed GroES oligomers are not required as functional cochaperonins. / Seale, Jeffrey W.; Chirgwin, John; Demeler, Borries; Horowitz, Paul M.

In: Journal of Protein Chemistry, Vol. 16, No. 7, 10.1997, p. 661-668.

Research output: Contribution to journalArticle

Seale, Jeffrey W. ; Chirgwin, John ; Demeler, Borries ; Horowitz, Paul M. / Preformed GroES oligomers are not required as functional cochaperonins. In: Journal of Protein Chemistry. 1997 ; Vol. 16, No. 7. pp. 661-668.
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