Introduction of a metal-dependent regulatory switch into an enzyme.
A cysteine has been introduced into the hydrophobic binding pocket of staphylococcal nuclease via oligonucleotide-directed mutagenesis. The L89C mutation does not significantly alter the catalytic activity or specificity of the nuclease yet provides a metal-dependent switch for regulating enzymatic activity. The L89C mutant can be inactivated by addition of mercuric or cupric salts and subsequently reactivated by addition of chelating agents. This work may provide a general strategy for regulating the catalytic activity of other enzymes or the binding affinity of proteins to DNA or other proteins.
Corey
D R
DR
Department of Chemistry, University of California, Berkeley 94270.
Schultz
P G
PG
D002614:Chelating Agents / Q000494:pharmacology; D003545:Cysteine; D004252:DNA Mutational Analysis; D005818:Genetic Engineering; D008670:Metals / Q000494:pharmacology*; D008836:Micrococcal Nuclease / Q000037:antagonists & inhibitors / Q000235:genetics*; D010662:Phenylmercuric Acetate / Q000494:pharmacology; D013329:Structure-Activity Relationship
D002614:Chelating Agents; D008670:Metals; D008836:Micrococcal Nuclease; D003545:Cysteine; D010662:Phenylmercuric Acetate