Redox reactions in the biosynthesis of thio-cofactors in bacteria

细菌硫代辅助因子生物合成中的氧化还原反应

• 批准号: 1716535
• 负责人: Patricia Dos Santos
• 金额: $ 68.07万
• 依托单位: Wake Forest University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1716535&HistoricalAwards=false
• 关键词: Redox; reactions; biosynthesis; thio; cofactors

Vitamins and other small molecules defined as cofactors that contain sulfur in their chemical structures are widely distributed in nature and essential for life on Earth. All known living organisms use at least a subset of these cofactors in various aspects of metabolism. While their importance in sustaining basic biochemical reactions is widely recognized, the mechanisms of sulfur activation, trafficking, and insertion into cofactors remain not fully understood. It is widely recognized, however, that the free amino acid cysteine is the source of sulfur for the biosynthesis of most sulfur-containing cofactors in bacteria, eukaryotes, and some species of archaea. A family of enzymes named cysteine desulfurases catalyze the first step in sulfur activation. This project will determine the effect of physiological reducing agents -specifically, thioredoxins and small molecules that contain sulfur (glutathione, bacillithiol, and mycothiol)- in biosynthetic reactions and identify specific protein interactions controlling the reactivity of cysteine desulfurases and their sulfur acceptors. This project will broaden the participation of underrepresented groups by including undergraduate researchers from Salem College and Guilford College, two smaller, local, primarily undergraduate institutions. Graduate students serving as co-mentors will gain improved communication skills in preparation as future independent teacher-scholars.The reaction catalyzed by cysteine desulfurases involves the formation and transfer of a persulfide intermediate to acceptor molecules involved in the biosynthesis of Fe-S clusters, thionucleosides in tRNA, rRNA and DNA, biotin, lipoic acid, molybdenum cofactor, and thiamine. Studying sulfur-transfer reactions in the biosynthesis of thio-cofactors is challenging. One of the major challenges is that sulfur-acceptor proteins are often shared across multiple biosynthetic pathways. This project will determine the involvement of physiological reductants in promoting redox reactions in the biosynthesis of thio-cofactors in bacteria by investigating different cysteine desulfurases that are involved in distinct pathways. The outcome of this study will provide fundamental knowledge regarding the role of redox agents in biochemical pathways involved in the transfer of sulfur and will establish evolutionary determinants controlling reactivity of biosynthetic enzymes. This project is supported jointly by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division in the Directorate for Biological Sciences and Chemistry for Life Processes Cluster of Chemistry Division in the Directorate for Mathematical and Physical Sciences.

维生素和其他小分子被定义为化学结构中含有硫的辅助因子，广泛分布在自然界中，对地球上的生命至关重要。所有已知的生物体在新陈代谢的各个方面都至少使用这些辅因子的一个子集。虽然它们在维持基本生化反应中的重要性已得到广泛认可，但硫的激活、运输和插入辅助因子的机制仍不完全清楚。然而，人们普遍认为游离氨基酸半胱氨酸是细菌、真核生物和某些古细菌物种中大多数含硫辅因子生物合成的硫源。名为半胱氨酸脱硫酶的酶家族催化硫活化的第一步。该项目将确定生理还原剂（特别是硫氧还蛋白和含硫小分子（谷胱甘肽、杆菌硫醇和菌硫醇））在生物合成反应中的作用，并确定控制半胱氨酸脱硫酶及其硫受体反应性的特定蛋白质相互作用。该项目将通过包括塞勒姆学院和吉尔福德学院这两个规模较小的本地本科院校的本科生研究人员来扩大代表性不足群体的参与。作为共同导师的研究生将获得更好的沟通技巧，为未来的独立教师学者做好准备。半胱氨酸脱硫酶催化的反应涉及过硫化物中间体的形成和转移到参与 Fe-S 簇、硫核苷生物合成的受体分子tRNA、rRNA 和 DNA、生物素、硫辛酸、钼辅因子和硫胺素。研究硫代辅助因子生物合成中的硫转移反应具有挑战性。主要挑战之一是硫受体蛋白通常在多种生物合成途径中共享。该项目将通过研究参与不同途径的不同半胱氨酸脱硫酶，确定生理还原剂在促进细菌硫代辅助因子生物合成中氧化还原反应中的参与。这项研究的结果将提供有关氧化还原剂在涉及硫转移的生化途径中的作用的基础知识，并将建立控制生物合成酶反应性的进化决定因素。该项目由生物科学局分子和细胞生物科学部分子生物物理学集群和数学和物理科学局化学司生命过程化学集群共同支持。

项目成果

Sulfur Availability Impacts Accumulation of the 2-Thiouridine tRNA Modification in Bacillus subtilis

硫的有效性影响枯草芽孢杆菌中 2-硫尿苷 tRNA 修饰的积累

• DOI: 10.1128/jb.00009-22
• 发表时间: 2022-05
• 期刊: Journal of Bacteriology
• 影响因子: 3.2
• 作者: Edwards, Ashley M.;Black, Katherine A.;Dos Santos, Patricia C.
• 通讯作者: Dos Santos, Patricia C.

Metallocluster transactions: dynamic protein interactions guide the biosynthesis of Fe–S clusters in bacteria

金属簇交易：动态蛋白质相互作用指导细菌中 Fe–S 簇的生物合成

• DOI: 10.1042/bst20180365
• 发表时间: 2018-12
• 期刊: Biochemical Society Transactions
• 影响因子: 3.9
• 作者: Zheng, Chenkang;Dos Santos, Patricia C.
• 通讯作者: Dos Santos, Patricia C.

Overview of physiological, biochemical, and regulatory aspects of nitrogen fixation in Azotobacter vinelandii

维氏固氮菌固氮的生理、生化和调控方面的概述

• DOI: 10.1080/10409238.2023.2181309
• 发表时间: 2022-11
• 期刊: Critical Reviews in Biochemistry and Molecular Biology
• 影响因子: 6.5
• 作者: Martin del Campo, Julia S.;Rigsbee, Jack;Bueno Batista, Marcelo;Mus, Florence;Rubio, Luis M.;Einsle, Oliver;Peters, John W.;Dixon, Ray;Dean, Dennis R.;Dos Santos, Patricia C.
• 通讯作者: Dos Santos, Patricia C.

Distribution of Nitrogen‐Fixation Genes in Prokaryotes Containing Alternative Nitrogenases

含替代固氮酶的原核生物中固氮基因的分布

• DOI: 10.1002/cbic.202000022
• 发表时间: 2020-04
• 期刊: ChemBioChem
• 影响因子: 3.2
• 作者: Addo, Maame A.;Dos Santos, Patricia C.
• 通讯作者: Dos Santos, Patricia C.

The Thioredoxin System Reduces Protein Persulfide Intermediates Formed during the Synthesis of Thio-Cofactors in Bacillus subtilis

硫氧还蛋白系统减少枯草芽孢杆菌硫代辅因子合成过程中形成的蛋白质过硫化物中间体

• DOI: 10.1021/acs.biochem.9b00045
• 发表时间: 2019-03
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Zheng, Chenkang;Guo, Selina;Tennant, William G.;Pradhan, Pradyumna K.;Black, Katherine A.;Dos Santos, Patricia C.
• 通讯作者: Dos Santos, Patricia C.