A mutant Cro protein, which bears the Ile-30----Leu substitution, is thermally unstable and degraded more rapidly than wild-type Cro in vivo. Using an antibody screen, we have isolated five different second site suppressor substitutions that reduce the proteolytic hypersensitivity of this mutant Cro protein. Two of the suppressor substitutions increase the thermal stability of Cro by 12 degrees C to 14 degrees C. These amino acid substitutions affect residues 16 and 26, which are substantially exposed to solvent in the crystal structure of wild-type Cro. Study holds ProTherm entries: 827, 828, 829, 830, 13437, 13438, 13439 Extra Details: enhanced stability; lambda Cro, genetic suppression;,intracellular proteolysis, antibody screen
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:16 p.m.
|Number of data points||11|
|Proteins||Regulatory protein cro ; Regulatory protein cro|
|Assays/Quantities/Protocols||Experimental Assay: ddG ; Experimental Assay: Tm ; Derived Quantity: dTm|
|Libraries||Mutations for sequence MEQRITLKDYAMRFGQTKTAKDLGVYQSAINKAIHAGRKIFLTINADGSVYAEEVKPFPSNKKTTA|