Ligand interaction scan: a general method for engineering ligand-sensitive protein alleles.
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).
Erster
Oran
O
Department of Biological Regulation, Weizmann Institute of Science, P.O. Box 26, Rehovot 76100, Israel.
Eisenstein
Miriam
M
Liscovitch
Mordechai
M
D023281:Genomics / Q000379:methods*; D007700:Kinetics; D008024:Ligands; D016254:Mutagenesis, Insertional; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000235:genetics*; D001618:beta-Lactamases / Q000235:genetics
D008024:Ligands; D011506:Proteins; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1