Mutant Proteins--Enzymes to Hydrolyze Toxic Organophosphates
Final rept. 15 Jun 1983-14 Jun 1989
CALIFORNIA INST OF TECH PASADENA DIV OF CHEMISTRY AND CHEMICAL ENGINEERING
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This work focuses on the creation of new catalytic activities by modification of existing enzymes. To this end, techniques for the synthesis of large genes have been developed together with a complementation system for the expression and correct folding of proteins that are normally self-processing but that have mutated so they can no longer process themselves. Concurrent expression of the pro-sequence and the mature protein sequence encoded on separate but compatible plasmids accomplishes this purpose the pro-sequence apparently performs the role of a chaperoning in this case. The enzymes studied are two serine hydrolases, alpha protease and B-lactamase. For a-lytic proteases the new activity sought was the ability to hydrolyze organophosphates rather than being irreversibly inhibited by these substances. For Beta lactamase we created a structural variant that had a dramatically different catalytic role on the normal beta lactam substrates.
- Medicine and Medical Research