Nucleotide-dependent oligomerization of ClpB from Escherichia coli

Michal Zolkiewski*, Martin Kessel, Ann Ginsburg, Michael R. Maurizi

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

75 Scopus citations

Abstract

Self-association of ClpB (a mixture of 95- and 80-kDa subunits) has been studied with gel filtration chromatography, analytical ultracentrifugation, and electron microscopy. Monomeric ClpB predominates at low protein concentration (0.07 mg/mL), while an oligomeric form is highly populated at >4 mg/mL. The oligomer formation is enhanced in the presence of 2 mM ATP or adenosine 5'-O-thiotriphosphate (ATPγS). In contrast, 2 mM ADP inhibits full oligomerization of ClpB. The apparent size of the ATP- or ATPγS-induced oligomer, as determined by gel filtration, sedimentation velocity and electron microscopy image averaging, and the molecular weight, as determined by sedimentation equilibrium, are consistent with those of a ClpB hexamer. These results indicate that the oligomerization reactions of ClpB are similar to those of other Hsp100 proteins.

Original languageEnglish
Pages (from-to)1899-1903
Number of pages5
JournalProtein Science
Volume8
Issue number9
DOIs
StatePublished - 1999
Externally publishedYes

Keywords

  • Electron microscopy
  • Heat-shock proteins
  • Nucleotide binding
  • Protein association
  • Sedimentation equilibrium
  • Sedimentation velocity

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