STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 细菌多组分单加氧酶（Bacterial multiplexor monooxygenase，BMPs）如甲烷单加氧酶（methane monooxygenase，MMO）、甲苯/邻二甲苯单加氧酶（toluene/o-xylene monooxygenase，ToMO）和苯酚羟化酶（phenol hydroxylase，PH）催化饱和烃和芳烃的区域选择性和对映选择性氧化，生成醇、酚、儿茶酚和环氧化物。需要调节和还原酶蛋白组分来实现BMM羟化酶中的催化作用，BMM羟化酶含有羧酸根桥接的非血红素二铁活性位点，在该位点处双氧活化的中间体形成并氧化烃底物。氙加压，基板/产品浸泡，和突变体形式的ToMO羟化酶和PH羟化酶和羟化酶调节复合物的晶体学表征的目的是探测基板访问途径，质子和电子转移机制，和结构-活性关系在Besides和二铁羧酸盐蛋白一般。这些数据将补充感兴趣的蛋白质的生物化学研究结果，并提供额外的信息与MMO系统进行比较，以阐明生物烃氧化的机制。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacterial multicomponent monooxygenases (BMMs) such as methane monooxygenase (MMO), toluene/o-xylene monooxygenase (ToMO), and phenol hydroxylase (PH) catalyze the regio- and enantioselective oxidation of saturated and aromatic hydrocarbons to yield alcohols, phenols, catechols, and epoxides. Regulatory and reductase protein components are required to effect catalysis in BMM hydroxylases, which contain carboxylate-bridged non-heme diiron active sites at which dioxygen-activated intermediates form and oxidize the hydrocarbon substrates. Crystallographic characterization of xenon-pressurized, substrate/product-soaked, and mutant forms of the ToMO hydroxylase and PH hydroxylase and hydroxylase-regulatory complex are sought to probe substrate access pathways, proton and electron transfer mechanisms, and structure-activity relationships in BMMs and diiron carboxylate protein in general. These data will complement biochemical findings on the proteins of interest and provide additional information for comparison with the MMO system in order to elucidate the mechanisms of biological hydrocarbon oxidation.