Site directed mutagenesis: a tool for enzyme mechanism dissection.
Site directed mutagenesis: a tool for enzyme mechanism dissection.
复制标题
定点诱变：酶机制剖析的工具。
DOI：
10.1016/0167-7799(90)90189-5
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发表时间：
1990
影响因子：
17.3
通讯作者：
Benkovic,SJ
中科院分区：
文献类型：
--
作者：
Wagner,CR;Benkovic,SJ
蛋白质工程已成为检测酶活性位点的主要手段，以鉴定和定量特定残基在配体结合、特异性和催化中的作用。定点诱变扩展了我们从X-射线晶体学获得的知识，并提供了惊人的证据，复杂的活性位点的几何形状是由蛋白质的基础设施的最大催化efficiency.Until最近，酶活性位点残基的鉴定和评估一直依赖于底物及其类似物的动力学行为的分析和蛋白质的化学修饰1，2。合成了具有潜在氢键、疏水相互作用或带电部分的缺失或添加的类似物。这种方法导致了强大的抑制剂的发现，在某些情况下是治疗上有用的3。
Protein engineering has become the principle means of examining the active site of an enzyme to identify and quantify the roles of specific residues in ligand binding, specificity and catalysis. Sitespecific mutagenesis has extended our knowledge gained from X-ray crystallography, and has provided striking proof that the intricate active-site geometry is supported by the remainder of the protein infrastructure for maximum catalytic efficiency.Until recently, identification and assessment of enzyme active site residues has relied on the analysis of the kinetic behavior of substrates and their analogs and on the chemical modification of proteins 1, 2. Analogs featuring deletion or addition of potential hydrogen bonds, hydrophobic interactions, or charged moieties were synthesized. This approach led to the discovery of powerful inhibitors which in some cases were therapeutically useful 3.