A water mediated electrostatic interaction gives thermal stability to the "tail" region of the GrpE protein from E. coli

Protein J. 2007 Jun;26(4):239-45. doi: 10.1007/s10930-006-9065-9.

Abstract

The GrpE protein from E. coli is a homodimer with an unusual structure of two long paired alpha-helices from each monomer interacting in a parallel arrangement to form a "tail" at the N-terminal end. Using site-directed mutagenesis, we show that there is a key electrostatic interaction involving R57 (mediated by a water molecule) that provides thermal stability to this "tail" region. The R57A mutant showed a drop in T (m) of 8.5 degrees C and a smaller DeltaH (u) (unfolding) compared to wild-type for the first unfolding transition, but no significant decrease in dimer stability as shown through equilibrium analytical ultracentrifugation studies. Another mutant (E94A) at the dimer interface showed a decrease in DeltaH (u )but no drop in T (m) for the second unfolding transition and a slight increase in dimer stability.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Circular Dichroism
  • Dimerization
  • Escherichia coli Proteins / chemistry*
  • Heat-Shock Proteins / chemistry*
  • Hot Temperature
  • Models, Molecular
  • Molecular Conformation
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Salts / chemistry
  • Static Electricity*
  • Temperature
  • Thermodynamics
  • Water / chemistry*

Substances

  • Escherichia coli Proteins
  • GrpE protein, E coli
  • Heat-Shock Proteins
  • Salts
  • Water