Phan Jason, Zdanov Alexander, Evdokimov Artem G, Tropea Joseph E, Peters Howard K, Kapust Rachel B, Li Mi, Wlodawer Alexander, Waugh David S
Macromolecular Crystallography Laboratory, Center for Cancer Research, NCI-Frederick, National Institutes of Health, Frederick, Maryland 21702-1201, USA.
J Biol Chem. 2002 Dec 27;277(52):50564-72. doi: 10.1074/jbc.M207224200. Epub 2002 Oct 10.
Because of its stringent sequence specificity, the 3C-type protease from tobacco etch virus (TEV) is frequently used to remove affinity tags from recombinant proteins. It is unclear, however, exactly how TEV protease recognizes its substrates with such high selectivity. The crystal structures of two TEV protease mutants, inactive C151A and autolysis-resistant S219D, have now been solved at 2.2- and 1.8-A resolution as complexes with a substrate and product peptide, respectively. The enzyme does not appear to have been perturbed by the mutations in either structure, and the modes of binding of the product and substrate are virtually identical. Analysis of the protein-ligand interactions helps to delineate the structural determinants of substrate specificity and provides guidance for reengineering the enzyme to further improve its utility for biotechnological applications.
由于其严格的序列特异性，烟草蚀纹病毒（TEV）的3C型蛋白酶常用于从重组蛋白中去除亲和标签。然而，目前尚不清楚TEV蛋白酶究竟是如何以如此高的选择性识别其底物的。现在，两个TEV蛋白酶突变体，即无活性的C151A和抗自溶的S219D，分别与底物肽和产物肽形成复合物，其晶体结构已在2.2埃和1.8埃分辨率下解析出来。在这两种结构中，酶似乎都没有受到突变的干扰，产物和底物的结合模式几乎相同。对蛋白质-配体相互作用的分析有助于描绘底物特异性的结构决定因素，并为改造该酶以进一步提高其在生物技术应用中的效用提供指导。
