Partial Opening of Cytochrome P450cam (CYP101A1) Is Driven by Allostery and Putidaredoxin Binding.
Partial Opening of Cytochrome P450cam (CYP101A1) Is Driven by Allostery and Putidaredoxin Binding.
复制标题
DOI：
10.1021/acs.biochem.1c00406
复制
发表时间：
2021-10-05
期刊：
影响因子：
2.9
通讯作者：
Paci E
中科院分区：
文献类型：
--
作者：
Skinner SP;Follmer AH;Ubbink M;Poulos TL;Houwing-Duistermaat JJ;Paci E
细胞色素P450 cam(CYP101A1)通过两个电子转移步骤的多步催化循环催化樟脑的区域和立体特异的5-外羟基化反应，绝对要求第二个电子由铁还蛋白-恶臭还蛋白(PDX)提供。在第二个电子转移步骤中，一旦樟脑结合到活性部位，底物进入通道关闭，P450cam是在PDX结合时打开，还是保持关闭，仍然存在争议。一个潜在的樟脑结合变构位点已经被确定，并被推测在PDX的结合中发挥作用。在这里，我们重新查看了顺磁核磁共振谱数据，并确定了解释数据的异质结构系综，提供了P450cam/PDX络合物在溶液中的完整表示，并协调了替代假设。变构樟脑结合位点总是存在的，樟脑与该结合位点的构象变化促进了PDX的结合。我们还确定了PDX结合的状态包括同时具有打开状态和关闭状态特征的结构集合。这些结果表明，在高樟脑浓度下，变构樟脑结合和PDX结合之间存在很好的平衡相互作用。
Cytochrome P450cam (CYP101A1) catalyzes the region- and stereo-specific 5-exo-hydroxylation of camphor via a multistep catalytic cycle that involves two-electron transfer steps, with an absolute requirement that the second electron be donated by the ferrodoxin, putidaredoxin (Pdx). Whether P450cam, once camphor has bound to the active site and the substrate entry channel has closed, opens up upon Pdx binding, during the second electron transfer step, or it remains closed is still a matter of debate. A potential allosteric site for camphor binding has been identified and postulated to play a role in the binding of Pdx. Here, we have revisited paramagnetic NMR spectroscopy data and determined a heterogeneous ensemble of structures that explains the data, provides a complete representation of the P450cam/Pdx complex in solution, and reconciles alternative hypotheses. The allosteric camphor binding site is always present, and the conformational changes induced by camphor binding to this site facilitates Pdx binding. We also determined that the state to which Pdx binds comprises an ensemble of structures that have features of both the open and closed state. These results demonstrate that there is a finely balanced interaction between allosteric camphor binding and the binding of Pdx at high camphor concentrations.
登录
查看更多内容
影响因子：
5.6
作者：
Asciutto EK;Pochapsky TC
通讯作者：
Pochapsky TC
影响因子：
5.5
作者：
Roe, Daniel R.;Cheatham, Thomas E., III
通讯作者：
Cheatham, Thomas E., III
影响因子：
5.6
作者：
POULOS, TL;FINZEL, BC;HOWARD, AJ
通讯作者：
HOWARD, AJ
DOI：
10.1073/pnas.1606474113
发表时间：
2016-08-02
影响因子：
11.1
作者：
Hollingsworth, Scott A.;Batabyal, Dipanwita;Poulos, Thomas L.
通讯作者：
Poulos, Thomas L.
Paramagpy：用于使用NMR光谱中测得的顺磁效应拟合磁敏感性张量的软件。
DOI：
10.5194/mr-1-1-2020
发表时间：
2020
期刊：
Magnetic resonance (Gottingen, Germany)
影响因子：
--
作者：
Orton HW;Huber T;Otting G
通讯作者：
Otting G