Normal cellular Ha ras p21 protein causes local disruption of bilayer phospholipid.
We have investigated the interactions of the p21 protein of c-Ha ras with its phospholipid environment. Gel filtration of detergent-"solubilized" p21 revealed that this preparation consisted of a mixture of multimolecular aggregates of protein and phospholipid and also a population of individual p21 molecules. Addition of 8 M urea to p21 preparations increased the solubility of the molecule in detergent solutions upon the removal of this denaturant. The progressive addition of the detergent cholate appeared to increase the efficiency of p21 preparations to bind GTP. This affinity for GTP was not removed even at high detergent concentrations, when delipification of the p21 was presumably effected. Modification of the composition of the phospholipid species surrounding the protein did not appear to alter its affinity for GTP. Electron spin resonance studies with membrane spin-labels indicated a perturbation of the bilayer extending to between 44 and 100 phospholipids surrounding the molecule. However, no evidence was found for any population of intimately bound phospholipid, which is seen as an annulus of about 30 lipids in transmembrane proteins such as Ca2+-ATPase. From these results we propose that the Ha ras p21 protein has the ability to associate directly with the membrane in a manner clearly discernible from that of a transmembrane protein.
Montgomery
G W
GW
Department of Chemistry, University of York, Heslington, England.
Jagger
B A
BA
Bailey
P D
PD
D002793:Cholic Acids; D002850:Chromatography, Gel; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D019204:GTP-Binding Proteins; D008051:Lipid Bilayers; D010743:Phospholipids / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D016283:Proto-Oncogene Proteins p21(ras)
D002793:Cholic Acids; D008051:Lipid Bilayers; D010743:Phospholipids; D011518:Proto-Oncogene Proteins; D019204:GTP-Binding Proteins; D016283:Proto-Oncogene Proteins p21(ras)