Conformational effects of nucleotide exchange in ras p21 proteins as studied by fluorescence spectroscopy
- 12 March 1990
- journal article
- Published by Wiley in FEBS Letters
- Vol. 262 (1) , 127-130
- https://doi.org/10.1016/0014-5793(90)80170-n
Abstract
The intrinsic fluorescence properties of the oncogene protein p21N-ras, p21H-ras and one of its transforming mutants, p21N-ras (Va1112), have been investigated. A mutant containing a single tryptophan at position 28 in p21H-ras (Trp28) has been specifically engineered to provide a probe of protein conformation on nucleotide binding. The proteins produced essentially similar circular dichroism spectra typical of alpha/beta proteins. A decrease in the intensity of the fluorescence emission spectrum due to tyrosine occurred on GDP/GTP nucleotide exchange in the native and mutant proteins. Selective excitation of the single tryptophan in p21 produced a decrease in fluorescence intensity which was accompanied by a blue shift in the wavelength of maximum emission on nucleotide exchange. A reduction in the residual Mg2+ ion concentration enhanced this effect.Keywords
This publication has 28 references indexed in Scilit:
- Structural differences between a ras oncogene protein and the normal proteinNature, 1989
- Conformational alterations detected by circular dichroism induced in the normal ras p21 protein by activating point mutations at position 12, 59, or 61European Journal of Biochemistry, 1988
- Spectroscopic and hydrodynamic studies reveal structural differences in normal and transforming H-ras gene productsBiochemistry, 1988
- The cytoplasmic protein GAP is implicated as the target for regulation by the ras gene productNature, 1988
- Cortical Flow in Animal CellsScience, 1988
- Homologies Between Signal Transducing G Proteins and ras Gene ProductsScience, 1984
- Activation of Ki-ras2 gene in human colon and lung carcinomas by two different point mutationsNature, 1983
- A point mutation is responsible for the acquisition of transforming properties by the T24 human bladder carcinoma oncogeneNature, 1982
- Mechanism of activation of a human oncogeneNature, 1982
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976