X-ray structural analysis of compensating mutations at the barnase-barstar interface

FEBS Lett. 1999 Jun 11;452(3):128-32. doi: 10.1016/s0014-5793(99)00621-3.

Abstract

The crystal structure of the barstar mutants (Y29P) and (Y29D, Y30W) as well as that of the complexes of barstar(Y29P) with wild-type barnase and barnase(H102K) have been determined. These barstar mutants compensate for the dramatic loss of barnase-barstar interaction energy caused by a single mutation of the barnase active site His-102 to a lysine. The latter introduces an uncompensated charge in the pocket at the surface of barstar where Lys-102 is located. The analysis of the structures suggests a mechanism for this compensation based on the solvation of the charge of Lys-102. Additional compensation occurs through the formation of a hydrogen bond.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Crystallography, X-Ray
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / metabolism
  • Histidine
  • Lysine
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Point Mutation
  • Protein Structure, Secondary
  • Ribonucleases / chemistry*
  • Ribonucleases / genetics
  • Ribonucleases / metabolism

Substances

  • Bacterial Proteins
  • Enzyme Inhibitors
  • barstar protein, Bacillus amyloliquefaciens
  • Histidine
  • Ribonucleases
  • Bacillus amyloliquefaciens ribonuclease
  • Lysine