Ligand interaction scan: a general method for engineering ligand-sensitive protein alleles
Oran Erster1, Miriam Eisenstein, Mordechai Liscovitch
1Department of Biological Regulation, Weizmann Institute of Science, P.O. Box 26, Rehovot 76100, Israel.
Abstract:
The ligand interaction scan (LIScan) method is a general procedure for engineering small molecule ligand-regulated forms of a protein that is complementary to other 'reverse' genetic and chemical-genetic methods for drug-target validation. It involves insertional mutagenesis by a chemical-genetic 'switch', comprising a genetically encoded peptide module that binds with high affinity to a small-molecule ligand. We demonstrated the method with TEM-1 beta-lactamase, using a tetracysteine hexapeptide insert and a biarsenical fluorescein ligand (FlAsH).
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