Protein dynamics in cytochrome P450 molecular recognition and substrate specificity using 2D IR vibrational echo spectroscopy.
Protein dynamics in cytochrome P450 molecular recognition and substrate specificity using 2D IR vibrational echo spectroscopy.
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DOI：
10.1021/ja109168h
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发表时间：
2011-03-23
影响因子：
15
通讯作者：
Fayer MD
中科院分区：
文献类型：
--
作者：
Thielges MC;Chung JK;Fayer MD
细胞色素（cyt）P450羟基化各种底物，这些底物的化学结构可能差异很大。这些酶在药物代谢和其他生物过程中的重要性促使人们研究使它们对不同类型分子具有活性的因素。蛋白质动力学与细胞色素P450底物特异性有关。在这里，2D红外振动回波光谱测量的动力学细胞色素P450凸轮从恶臭假单胞菌在快速的时间尺度上使用CO结合在活性位点作为振动探针。无底物的酶和酶绑定到其天然底物，樟脑，和一系列相关的基板进行了研究，以阐明在细胞色素P450CAM的分子识别动力学的作用，并描绘如何运动可能有助于羟基化特异性。在无底物的细胞色素P450cam中，三种构象状态被填充，并且构象状态内的结构波动相对缓慢。底物结合选择性地稳定一种构象状态，并且动力学变得更快。在所观察到的动力学与底物的羟基化的特异性，结合亲和力，和底物的分子体积的相关性表明，在数百ps的时间尺度上的运动有助于细胞色素P450cam对不同底物的活性的变化。
Cytochrome (cyt) P450s hydroxylate a variety of substrates that can differ widely in their chemical structure. The importance of these enzymes in drug metabolism and other biological processes has motivated the study of the factors that enable their activity on diverse classes of molecules. Protein dynamics have been implicated in cyt P450 substrate specificity. Here, 2D IR vibrational echo spectroscopy is employed to measure the dynamics of cyt P450cam from Pseudomonas putida on fast timescales using CO bound at the active site as a vibrational probe. The substrate-free enzyme and the enzyme bound to both its natural substrate, camphor, and a series of related substrates are investigated to explicate the role of dynamics in molecular recognition in cyt P450cam and to delineate how the motions may contribute to hydroxylation specificity. In substrate-free cyt P450cam three conformational states are populated, and the structural fluctuations within a conformational state are relatively slow. Substrate binding selectively stabilizes one conformational state, and the dynamics become faster. Correlations in the observed dynamics with the specificity of hydroxylation of the substrates, the binding affinity, and the substrates’ molecular volume suggest that motions on the hundreds of ps timescale contribute to the variation in activity of cyt P450cam towards different substrates.
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影响因子：
3.3
作者：
Finkelstein, IJ;Goj, A;Fayer, MD
通讯作者：
Fayer, MD
影响因子：
4.4
作者：
Eyring, H
通讯作者：
Eyring, H
影响因子：
3.9
作者：
Hamuro, Yoshitomo;Molnar, Kathleen S.;Pochapsky, Thomas C.
通讯作者：
Pochapsky, Thomas C.
影响因子：
15
作者：
HARRIS, D;LOEW, G
通讯作者：
LOEW, G
DOI：
10.1073/pnas.0709760104
发表时间：
2007-12-04
影响因子：
11.1
作者：
Ishikawa, Haruto;Kim, Seongheun;Fayer, Michael D.
通讯作者：
Fayer, Michael D.