Ligand interaction scan: a general method for engineering ligand-sensitive protein alleles.

Erster, Oran; Eisenstein, Miriam; Liscovitch, Mordechai · Nat Methods · 2007

basic_science · Level V

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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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