疏水残基在二氢叶酸还原酶的结合与催化中的重要性。
Importance of a hydrophobic residue in binding and catalysis by dihydrofolate reductase.
作者信息
Mayer R J, Chen J T, Taira K, Fierke C A, Benkovic S J
出版信息
Proc Natl Acad Sci U S A. 1986 Oct;83(20):7718-20. doi: 10.1073/pnas.83.20.7718.
Abstract
A conserved residue at the dihydrofolate binding site of dihydrofolate reductase (EC 1.5.1.3), leucine-54, was replaced with glycine to ascertain the role of this hydrophobic amino acid. The effect of the mutation is both to increase the dissociation rate of dihydrofolate and decrease the rate of hydride transfer thus changing the rate-limiting step in catalysis from product loss (leucine-54) to hydride transfer (glycine-54). The total stabilization by leucine-54 of the transition state for hydride transfer is ca. 10(4)-fold (delta delta G approximately 5.4 kcal/mol) at subsaturating dihydrofolate levels relative to free enzyme despite its location some 10 A from the site of chemical reaction.
摘要
二氢叶酸还原酶（EC 1.5.1.3）二氢叶酸结合位点的一个保守残基亮氨酸-54被甘氨酸取代，以确定这种疏水氨基酸的作用。该突变的作用是既增加二氢叶酸的解离速率，又降低氢化物转移速率，从而将催化中的限速步骤从产物损失（亮氨酸-54）转变为氢化物转移（甘氨酸-54）。尽管亮氨酸-54距离化学反应位点约10埃，但在亚饱和二氢叶酸水平下，相对于游离酶，亮氨酸-54对氢化物转移过渡态的总稳定作用约为10^4倍（ΔΔG约为5.4千卡/摩尔）。
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