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Science 5 July 1996:
Vol. 273. no. 5271, pp. 115 - 117
DOI: 10.1126/science.273.5271.115
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Reports

Formation of a Transition-State Analog of the Ras GTPase Reaction by Ras·GDP, Tetrafluoroaluminate, and GTPase-Activating Proteins

Rohit Mittal, * Mohammad Reza Ahmadian, * Roger S. Goody, Alfred Wittinghofer dagger

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 (AlF4-). However, the product of the proto-oncogene ras in its guanosine diphosphate (GDP)-bound form interacted with AlF4- 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.

R. Mittal, M. R. Ahmadian, A. Wittinghofer, Abteilung Strukturelle Biologie, Max-Planck-Institut für Molekulare Physiologie, Rheinlanddamm 201, 44139 Dortmund, Germany.
R. S. Goody, Abteilung Physikalische Biochemie, Max-Planck-Institut für Molekulare Physiologie, Rheinlanddamm 201, 44139 Dortmund, Germany.
* These authors contributed equally to this work.
dagger To whom correspondence should be addressed.


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