Binding of a chaperonin to the folding intermediates of lactate dehydrogenase

I G Badcoe, C J Smith, S Wood, D J Halsall, J J Holbrook, P Lund, A R Clarke

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138 Citations (Scopus)

Abstract

When Bacillus stearothermophilus LDH dimer is incubated with increasing concentrations of the denaturant guanidinium chloride, three distinct unfolded states of the molecule are observed at equilibrium [Smith, C. J., et al. (1991) Biochemistry 30, 1028-1036]. The kinetics of LDH refolding are consistent with an unbranched progression through these states. The Escherichia coli chaperonin, GroEL, binds with high affinity to the completely denatured form and more weakly to the earliest folding intermediate, thus retarding the refolding process. A later structurally defined folding intermediate, corresponding to a molten globule form, is not bound by GroEL; neither is the inactive monomer. The complex between GroEL and denatured LDH is destabilized by the binding of magnesium/ATP (Mg/ATP) or by the nonhydrolyzable analogue adenylyl imidodiphosphate (AMP-PNP). From our initial kinetic data, we propose that GroEL exists in two interconvertible forms, one of which is stabilized by the binding of Mg/ATP but associates weakly with the unfolded protein. The other is destabilized by Mg/ATP and associates strongly with unfolded LDH. The relevance of these findings to the role of GroEL in vivo is discussed.
Original languageEnglish
Pages (from-to)9195-200
Number of pages6
JournalBiochemistry
Volume30
Issue number38
Publication statusPublished - 24 Sep 1991

Keywords

  • Microscopy, Fluorescence
  • L-Lactate Dehydrogenase
  • Bacterial Proteins
  • Adenine Nucleotides
  • Chaperonin 60
  • Chaperonins
  • Protein Denaturation
  • Proteins
  • Geobacillus stearothermophilus
  • Protein Binding
  • Protein Conformation
  • Heat-Shock Proteins

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