Nature Methods
- 4, 393 - 395 (2007)
Published online: 22 April 2007; | doi:10.1038/nmeth1046
Ligand interaction scan: a general method for engineering ligand-sensitive protein allelesOran Erster1, Miriam Eisenstein2 & Mordechai Liscovitch11
Department of Biological Regulation, Weizmann Institute of Science, P.O. Box 26, Rehovot 76100, Israel. 2
Department of Chemical Research Support, Weizmann Institute of Science, P.O. Box 26, Rehovot 76100, Israel.
Correspondence should be addressed to Mordechai Liscovitch moti.liscovitch@weizmann.ac.il 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  -lactamase, using a tetracysteine hexapeptide insert and a biarsenical fluorescein ligand (FlAsH).
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