Characterization of pmoD

 描述（由申请人提供）：甲烷是一种导致全球变暖的常见温室气体。虽然合成上难以将甲烷化学转化为 甲烷氧化菌是一种对环境危害较小的物质，它已经开发出一种代谢甲烷的有效方法。在该过程的第一步中，甲烷氧化菌使用一种称为颗粒甲烷单加氧酶（pMMO）的铜依赖性酶复合物将甲烷转化为甲醇，但这是如何在生物化学上发生的，以及pMMO是如何被调节的还没有很好的了解。该提案的主要目标是表征一种名为pmoD的蛋白质，该蛋白质也可能参与甲烷转化，并可能以某种方式调节pMMO。在这项工作的一个组成部分中，将使用pmoD敲除菌株研究pmoD的表型。第二方面的工作将集中在结构，生物化学和光谱表征的pmoD。第三个目标是通过确定pmoD是否可以与参与铜代谢和甲烷转化的其他蛋白质相互作用以及是否可以调节这些过程来解决pmoD的功能。总之，体内研究、结构表征和生化分析的这种组合预期提供pMoD及其在甲烷代谢和铜调节中的作用的全面描述。从这些研究中获得的见解有可能大大提高目前对生物甲烷氧化的理解。生物化学分析有望提供一个全面的描述pmoD及其在甲烷代谢和铜调节的作用。从这些研究中获得的见解有可能大大提高目前对生物甲烷氧化的理解。

 DESCRIPTION (provided by applicant): Methane is a prevalent greenhouse gas that contributes to global warming. While synthetically it is difficult to chemically convert methane to less environmentally harmful substances, methanotrophic bacteria have developed an efficient way to metabolize methane. In the first step of the process, methanotrophs use a copper dependent enzyme complex called particulate methane monooxygenase (pMMO) to convert methane to methanol, but precisely how this occurs biochemically and how pMMO may be regulated are not well understood. The primary goal of this proposal is to characterize a protein called pmoD that is also likely involved in methane conversion and may somehow regulate pMMO. In one component of this work the phenotype of pmoD will be studied using a pmoD knockout strain. A second aspect of the work will focus on structural, biochemical, and spectroscopic characterization of pmoD. The third aim addresses the function of pmoD by determining whether it can interact with other proteins involved in copper metabolism and methane conversion and whether it can regulate these processes. Together, this combination of in'vivo studies, structural characterization, and biochemical analysis is expected to provide a comprehensive description of pmoD and its roles in methane metabolism and copper regulation. The insights gained from these studies have the potential to greatly enhance the current understanding of biological methane oxidation. Biochemical analysis is expected to provide a comprehensive description of pmoD and its roles in methane metabolism and copper regulation. The insights gained from these studies have the potential to greatly enhance the current understanding of biological methane oxidation.