Ligand interaction scan: a general method for engineering ligand-sensitive protein alleles
Ligand interaction scan: a general method for engineering ligand-sensitive protein alleles
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DOI：
10.1038/nmeth1046
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发表时间：
2007-05-01
期刊：
影响因子：
48
通讯作者：
Liscovitch, Mordechai
中科院分区：
文献类型：
--
作者：
Erster, Oran;Eisenstein, Miriam;Liscovitch, Mordechai
配体相互作用扫描（LIScan）方法是用于工程化小分子配体调节形式的蛋白质的通用程序，其与用于药物靶标验证的其它“反向”遗传和化学遗传方法互补。它涉及通过化学-遗传“开关”的插入诱变，所述开关包含以高亲和力结合小分子配体的遗传编码的肽模块。我们证明了TEM-1 β-内酰胺酶的方法，使用四半胱氨酸六肽插入和双砷荧光素配体（FlAsH）。
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).