Browner M F, Hackos D, Fletterick R
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448, USA.
Nat Struct Biol. 1994 May;1(5):327-33. doi: 10.1038/nsb0594-327.
Activation of protein function through phosphorylation can be mimicked by the engineering of specific metal binding sites. The addition of two histidine residues to glycogen phosphorylase allows enzymatic activation by transition metals in a cooperative and allosteric manner. Crystal structures of the metallo-enzyme have been determined and show that the structural transition induced upon metal binding (Ni2+) is, in part, analogous to the mode of activation of the native enzyme. The designed metal activation site allows assignment of the structural changes which trigger activation in this allosteric enzyme and, further, provide insight into the evolutionary development of multiple activation sites.
通过特定金属结合位点的工程设计，可以模拟磷酸化对蛋白质功能的激活作用。在糖原磷酸化酶上添加两个组氨酸残基，可使过渡金属以协同和变构的方式激活酶活性。已确定该金属酶的晶体结构，结果表明金属结合（Ni2+）时诱导的结构转变部分类似于天然酶的激活模式。设计的金属激活位点能够确定在这种变构酶中触发激活的结构变化，进而深入了解多个激活位点的进化发展。
