Biochemical and crystallographic characterization of a complex of c-Ha-ras p21 and caged GTP with flash photolysis.
- 1 October 1989
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 86 (20), 7687-7690
- https://doi.org/10.1073/pnas.86.20.7687
Abstract
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(p21''c) and the corresponding region from an oncogenic mutant form of the protein in which glycine at position 12 has been replaced by valine [p21''(G12V)] have been crystallized with P3-1-(2-nitro)phenylethylguanosine 5''-O-triphosphate (caged GTP) at their active sites.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 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 obtianed 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.This publication has 15 references indexed in Scilit:
- C-terminal Truncation of p21H Preserves Crucial Kinetic and Structural PropertiesJournal of Biological Chemistry, 1989
- Crystallization and preliminary X-ray analysis of the human c-H-ras-oncogene product p21 complexed with GTP analoguesJournal of Molecular Biology, 1989
- Millisecond X-ray diffraction and the first electron density map from Laue photographs of a protein crystalNature, 1987
- ras GENESAnnual Review of Biochemistry, 1987
- Preparation and characterization of nucleotide-free and metal ion-free p21 “apoprotein”.Journal of Biological Chemistry, 1987
- Characterisation of the metal‐ion–GDP complex at the active sites of transforming and nontransforming p21 proteins by observation of the 17O‐Mn superhyperfine coupling and by kinetic methodsEuropean Journal of Biochemistry, 1986
- Time-Resolved Structural Studies on Insect Flight Muscle after Photolysis of Caged-ATPBiophysical Journal, 1986
- A new approach to time-resolved studies of ATP-requiring biological systems; laser flash photolysis of caged ATP.Proceedings of the National Academy of Sciences, 1980
- The Binding Constant of ATP to Myosin S1 FragmentEuropean Journal of Biochemistry, 1977
- Conversion of Mono- and Oligodeoxyribonucleotides to 5'-Triphosphates1Journal of the American Chemical Society, 1965