Open AccessBiochemical and crystallographic characterization of a complex of c-Ha-ras p21 and caged GTP with flash photolysis.
Author(s) -
Ilme Schlichting,
Gert Rapp,
Jacob John,
Alfred Wittinghofer,
E.F. Pai,
Roger S. Goody
Publication year - 1989
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.86.20.7687
Subject(s) - gtp' , flash photolysis , chemistry , photodissociation , crystallography , hydrolysis , mutant , guanosine triphosphate , glycine , stereochemistry , biochemistry , enzyme , photochemistry , amino acid , kinetics , reaction rate constant , quantum mechanics , gene , physics
The GTP binding domain of the c-Ha-ras protooncogene product (p21'c) and the corresponding region from an oncogenic mutant form of the protein in which glycine at position 12 has been replaced by valine [p21'(G12V)] have been crystallized with P3-1-(2-nitro)phenylethylguanosine 5'-O-triphosphate (caged GTP) at their active sites. The crystals give x-ray diffraction patterns to a resolution of better than 0.3 nm. Photolysis can be achieved in the crystal, after which GTP hydrolysis takes place at the rate expected from solution studies. Complete x-ray data sets have been obtained for the starting caged-GTP state and the final GDP state after photolysis and hydrolysis, demonstrating the feasibility of time-resolved structural investigations of the process of GTP hydrolysis.