| Popis: |
The Kras gene encodes a small GTPase that is involved in pathways regulating cell growth and proliferation. Bound to GTP, Kras sends growth signals until the GTP is hydrolyzed. Hydrolysis is catalyzed by GTPase Activating Protein (GAP). Activating mutations in Kras are highly correlated to cancer development due to uncontrolled growth. Any mutation in the 12th amino acid of Kras (glycine) cause steric alterations in the catalytic site of GAP that prevent catalysis of GTP hydrolysis. Kras is a widely-studied signaling protein, and currently there are no clinical inhibitors. This project will attempt to redesign the catalytic site of GAP to circumvent the steric interference caused by G12 mutations, thus allowing hydrolysis to occur and signaling to be turned off. Recombinant proteins were purified from E. coli, and GAP mutants were screened for catalytic activity using a colorimetric GTPase assay. |
