Formation of a transition-state analog of the Ras GTPase reaction by Ras-GDP, tetrafluoroaluminate, and GTPase-activating proteins

Science. 1996 Jul 5;273(5271):115-7. doi: 10.1126/science.273.5271.115.

Abstract

Unlike the alpha subunits of heterotrimeric guanosine triphosphate (GTP)-binding proteins, Ras-related GTP-binding proteins have hitherto been considered not to bind or become activated by tetrafluoroaluminate (AIF4-). However, the product of the proto-oncogene ras in its guanosine diphosphate (GDP)-bound form interacted with AIF4 - in the presence of stoichiometric amounts of either of the guanosine triphosphatase (GTPase)-activating proteins (GAPs) p120GAP and neurofibromin. Neither oncogenic Ras nor a GAP mutant without catalytic activity produced such a complex. Together with the finding that the Ras-binding domain of the protein kinase c-Raf, whose binding site on Ras overlaps that of the GAPs, did not induce formation of such a complex, this result suggests that GAP and neurofibromin stabilize the transition state of the GTPase reaction of Ras.

MeSH terms

  • Aluminum Compounds / metabolism*
  • Amino Acid Sequence
  • Fluorides / metabolism*
  • GTP Phosphohydrolases / metabolism*
  • GTPase-Activating Proteins
  • Guanosine Diphosphate / metabolism
  • Molecular Sequence Data
  • Mutagenesis
  • Neurofibromin 1
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism*
  • Spectrometry, Fluorescence
  • ras GTPase-Activating Proteins
  • ras Proteins / metabolism*

Substances

  • Aluminum Compounds
  • GTPase-Activating Proteins
  • Neurofibromin 1
  • Proteins
  • ras GTPase-Activating Proteins
  • Guanosine Diphosphate
  • tetrafluoroaluminate
  • GTP Phosphohydrolases
  • ras Proteins
  • Fluorides