Atomic structures and reaction mechanisms of proteins involved in transfer and insertion of molybdenum cofactor and heme

参与钼辅因子和血红素转移和插入的蛋白质的原子结构和反应机制

• 批准号: 131891027
• 负责人: Professor Dr. Dirk Heinz
• 金额: --
• 依托单位: Helmholtz-Zentrum für Infektionsforschung (HZI)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/131891027?language=en
• 关键词: Atomic; structures; reaction; mechanisms; proteins

This project is a collaborative project interacting with four other projects within the Forschergruppe. The aim is to crystallize the following proteins and solve their atomic structures in order to decipher their hitherto unknown reaction mechanisms: (1) The molybdenum cofactor (Moco)-carrying ABA3 sulfurase of Arabidopsis alone and in complex with its target enzyme xanthine dehdrogenase and/or aldehyde oxidase. Here we want to unravel the mechanism by which ABA3 achieves the insertion of Moco and the sulfuration of the Mo-center as required by xanthine dehydrogenase and aldehyde oxidase. (2) The Arabidopsis thaliana Moco-donor protein Cnx1 in complex with its substrate MPT-AMP. Structure-guided mutagenesis will provide insight into MPT-AMP hydrolysis and will shed light onto the mechanism how Moco is presented by the Cnx1 Moco-donor platform. (3) The heme-binding protein HemW from Escherichia coli with and without bound heme. Since the [4Fe-4S] cluster and S-adenoslymethionine containing protein is dimerizing upon heme binding and subsequently transferring electrons, involved structural changes and possible catalytic principle are of interest. (4) The heme d1 insertion protein NirN in complex with heme d1 and in complex with its target protein NirS. Here we want to analyze how heme d1 binding in NirN differs from that in NirS and how the two proteins interact with each other for heme d1 transfer from NirN to NirS.

这个项目是一个协作项目，与Forschergruppe内的其他四个项目互动。本研究的目的是对以下蛋白质进行结晶并解析它们的原子结构，以破解它们迄今未知的反应机制：(1)含钼辅因子(MOCO)的拟南芥ABA3硫酶及其靶酶黄嘌呤脱氢酶和/或醛氧化酶的复合体。在这里，我们想要揭示ABA3实现黄嘌呤脱氢酶和醛氧化酶所需的MoCO插入和Mo中心硫化的机制。(2)拟南芥模拟供体蛋白Cnx1与其底物MPT-AMP的复合体。结构导向突变将为MPT-AMP的水解提供洞察力，并将揭示Cnx1 Moco供体平台如何呈现Moco的机制。(3)含和不含结合血红素的大肠杆菌中的血红素结合蛋白hemW。由于含有[4Fe-4S]簇和S-腺苷蛋氨酸的蛋白质在血红素结合时发生二聚，随后转移电子，涉及的结构变化和可能的催化原理是感兴趣的。(4)血红素D1插入蛋白Nirn与血红素D1及其靶蛋白NIRS的复合体。在这里，我们想分析Nirn和NIRS中的血红素D1结合有何不同，以及这两种蛋白质在血红素D1从Nirn转移到NIRS的过程中是如何相互作用的。