Structure of the carboxyl terminus of the RAS gene-encoded P21 proteins.

Publisher Website
Google Scholar
Abstract
The three-dimensional structures of the carboxyl-terminal regions of the P21 protein products of the human Harvey (Ha), Kirsten (KiA and KiB), and neuroblastoma (N) RAS oncogenes and various mutants have been determined by using conformational energy analysis. The carboxyl-terminal regin of P21 has been strongly implicated in the binding of the protein to the inner surface of the plasma membrane without which the protein is inactive. The only invariant residue in this region is Cys-186, which is necessary for the post-translational addition of palmitic acid. The surrounding sequences of the active native proteins differ considerably. Nevertheless, certain amino acid substitutions in this region are known to eliminate membane binding and protein activity, suggesting that there is a conserved common structural feature in this region in the native proteins that is disrupted in the mutant proteins. Conformational energy analysis shows that the four native P21 proteins have a common structure in the form of an .alpha.-helix for the terminal pentapeptide. A mutant, pBW277, that fails to bind to the membrane and is inactive cannot adopt an .alpha.-helical structure in this region because of a proline at position 188. Another mutant, pBW766, that retains membane binding and activity, on the other hand, retains the preference for an .alpha.-helical conformation in the terminal pentapeptide. These findings suggest that, despite various amino acid sequences in this region, the carboxylterminal pentapeptides of the P21 proteins form a distinctive structural domain that must have an .alpha.-helical structure for membrane binding and intracellular activity.