Probing the inhibitor selectivity pocket of human 20α-hydroxysteroid dehydrogenase (AKR1C1) with X-ray crystallography and site-directed mutagenesis
Probing the inhibitor selectivity pocket of human 20α-hydroxysteroid dehydrogenase (AKR1C1) with X-ray crystallography and site-directed mutagenesis
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DOI：
10.1016/j.bmcl.2011.01.076
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发表时间：
2011-04-15
影响因子：
2.7
通讯作者：
Hara, Akira
中科院分区：
文献类型：
--
作者：
El-Kabbani, Ossama;Dhagat, Urmi;Hara, Akira
人类20α-羟基类固醇脱氢酶(AKR1C1)是一个重要的药物靶点，因为它在肺癌、子宫内膜癌、早产和神经元疾病的发生中发挥作用。报道了AKR1C1与活性中心结合的第一个结构阻滞剂3-氯-5-苯基水杨酸(K(I)=0.86 nm)的晶体结构。3-氯-5-苯基水杨酸与AKR1C1的结合导致了Phe311侧链的构象变化，以适应抑制剂5-位上的大体积苯环取代基。通过定点突变进一步研究了非保守残基Leu54、Leu306、Leu308和Phe311对结合的贡献，并测定了突变对K(I)值的影响。与野生型酶相比，Leu54Val和Leu306Ala突变分别导致K(I)值增加6倍和81倍，而其余突变几乎没有影响。(C)爱思唯尔有限公司出版的2011年。
Human 20 alpha-hydroxysteroid dehydrogenase (AKR1C1) is an important drug target due to its role in the development of lung and endometrial cancers, premature birth and neuronal disorders. We report the crystal structure of AKR1C1 complexed with the first structure-based designed inhibitor 3-chloro-5-phenylsalicylic acid (K(i) = 0.86 nM) bound in the active site. The binding of 3-chloro-5-phenylsalicylic acid to AKR1C1 resulted in a conformational change in the side chain of Phe311 to accommodate the bulky phenyl ring substituent at the 5-position of the inhibitor. The contributions of the nonconserved residues Leu54, Leu306, Leu308 and Phe311 to the binding were further investigated by site-directed mutagenesis, and the effects of the mutations on the K(i) value were determined. The Leu54Val and Leu306Ala mutations resulted in 6- and 81-fold increases, respectively, in K(i) values compared to the wild-type enzyme, while the remaining mutations had little or no effects. (C) 2011 Published by Elsevier Ltd.