Transient and Stable Macromolecular Complexes Formed by Denitrifying Enzymes

反硝化酶形成的瞬时和稳定的大分子复合物

• 批准号: BB/L006960/1
• 负责人: Samar Hasnain
• 金额: $ 74.45万
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL006960%2F1
• 关键词: Transient; Stable; Macromolecular; Complexes; Formed

Redox proteins, including metalloproteins, form a large portion of the protein kingdom. Metalloproteins themselves form ~ 30% of a genome. These contain metal ions either as a single atom or as part of a cluster and play a variety of life sustaining roles in the bacterial, plant and animal kingdoms. Many enzymes exploit the oxidation states of metals to perform redox cycling. Fundamental biological processes in which metalloproteins participate include electron storage and transfer, dioxygen binding, storage and activation, and substrate transport, catalysis and activation. In many metalloenzymes such as cytochrome c oxidase (essential for mammalian life through respiratory requirements), nitrogenases and nitrite reductases (essential in view of their central position in the nitrogen cycle), hydrogenases (producers of molecular hydrogen - a candidate for a future alternative energy source), catalysis involves the controlled delivery of electrons and protons to the active site where substrate is utilised. The proposal build on close collaboration between the applicants and the RIKEN group (Japan) where they collectively have made major contributions in the field of denitrification and have provided significant advances in our understanding of complex processes that are involved in biological mechanisms of metalloenzymes. Our combined approaches have allowed us to build detailed three dimensional structural movies of catalysis in the crystalline state, thereby providing detailed insight into enzyme reaction chemistry. Our recent determination of structures for the membrane proteins, nitric oxide reducatses (Tosha & Shiro, RIKEN) together with the atomic resolution structure of the tethered cytochrome-Cu1-Cu2 nitrite reductase (Antonyuk, Eady & Hasnain, UoL) puts us in a very strong position to establish an integrated structural-mechanistic biology programme. This programme is aimed at understanding complex mechanisms of redox control, regulation and communication in globally important biological systems. General principles emerging from these studies will underpin our understanding of the control of redox processes in biology and protection against toxic chemical intermediates like nitric oxide. New methods and approaches developed in this programme (e.g. development of laboratory-based size-exclusion chromatography-small angle X-ray scattering with dynamic light scattering (SEC-SAXS-DLS) for studying protein complexes) will have broad relevance to our capabilities for studying protein complexes. These new capabilities and the scientific outcome will have significant impact on structural-mechanistic biology and keep the UK at the forefront of global effort in this important field.

氧化还原蛋白，包括金属蛋白，构成了蛋白质王国的很大一部分。金属蛋白本身约占基因组的30%。它们含有金属离子，或作为单个原子，或作为簇的一部分，在细菌、植物和动物王国中起着各种维持生命的作用。许多酶利用金属的氧化态来进行氧化还原循环。金属蛋白参与的基本生物过程包括电子储存和转移、二氧结合、储存和激活、底物运输、催化和激活。在许多金属酶中，如细胞色素c氧化酶（哺乳动物生命通过呼吸需求必不可少），氮酶和亚硝酸盐还原酶（鉴于它们在氮循环中的中心位置而必不可少），氢化酶（分子氢的生产者-未来替代能源的候选物），催化包括电子和质子的受控传递到利用底物的活性位点。该提案建立在申请人和RIKEN集团（日本）之间的密切合作基础上，他们共同在反硝化领域做出了重大贡献，并为我们对涉及金属酶生物机制的复杂过程的理解提供了重大进展。我们的联合方法使我们能够建立晶体状态下催化的详细三维结构电影，从而为酶反应化学提供详细的见解。我们最近对膜蛋白结构的测定，一氧化氮还原（Tosha & Shiro， RIKEN）以及拴系细胞色素- cu1 - cu2亚硝酸盐还原酶的原子分辨率结构（Antonyuk, Eady & Hasnain, UoL）使我们处于非常有利的地位，以建立一个集成的结构-机制生物学程序。该计划旨在了解全球重要生物系统中氧化还原控制、调节和交流的复杂机制。从这些研究中得出的一般原则将巩固我们对生物学中氧化还原过程的控制和对有毒化学中间体（如一氧化氮）的保护的理解。在这个项目中开发的新方法和方法（例如，用于研究蛋白质复合物的基于实验室的尺寸排除色谱-小角度x射线散射与动态光散射（SEC-SAXS-DLS）的发展）将与我们研究蛋白质复合物的能力具有广泛的相关性。这些新的能力和科学成果将对结构机制生物学产生重大影响，并使英国在这一重要领域保持全球领先地位。

项目成果

An unprecedented dioxygen species revealed by serial femtosecond rotation crystallography in copper nitrite reductase.

• DOI: 10.1107/s2052252517016128
• 发表时间: 2018-01-01
• 期刊: IUCrJ
• 影响因子: 3.9
• 作者: Halsted TP;Yamashita K;Hirata K;Ago H;Ueno G;Tosha T;Eady RR;Antonyuk SV;Yamamoto M;Hasnain SS
• 通讯作者: Hasnain SS

Sub-atomic resolution X-ray crystallography and neutron crystallography: promise, challenges and potential.

• DOI: 10.1107/s2052252515011239
• 发表时间: 2015-07-01
• 期刊: IUCrJ
• 影响因子: 3.9
• 作者: Blakeley MP;Hasnain SS;Antonyuk SV
• 通讯作者: Antonyuk SV

Identification of a tyrosine switch in copper-haem nitrite reductases.

• DOI: 10.1107/s2052252518008242
• 发表时间: 2018-07-01
• 期刊: IUCrJ
• 影响因子: 3.9
• 作者: Dong J;Sasaki D;Eady RR;Antonyuk SV;Hasnain SS
• 通讯作者: Hasnain SS

Characterization of the quinol-dependent nitric oxide reductase from the pathogen Neisseria meningitidis, an electrogenic enzyme.

• DOI: 10.1038/s41598-018-21804-0
• 发表时间: 2018-02-26
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Gonska N;Young D;Yuki R;Okamoto T;Hisano T;Antonyuk S;Hasnain SS;Muramoto K;Shiro Y;Tosha T;Ädelroth P
• 通讯作者: Ädelroth P