权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Studies of metal-containing and metal-less EutT adenosyltransferases

含金属和无金属EutT腺苷基转移酶的研究

基本信息

批准号：
10058537
负责人：
Flavia Gisela Costa
金额：
$ 4.24万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2020-12-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10058537
关键词：
Acetaldehyde Acetyl Coenzyme A Address Anabolism Animals Area Biochemical Biochemistry Biological Biophysics Catabolism Catalysis Cell physiology Cells Cellular biology Chemistry Coenzyme A Coenzymes Collaborations Complex Complex Analysis Corrinoids Crystallography Development Environment Enzymatic Biochemistry Enzymes Ethanolamine Ammonia-Lyase Ethanolamines Eukaryota Family Future Genetic Goals Health Homologous Gene Human Intestines Knowledge Laboratories Life Listeria monocytogenes Location Mediating Metabolic Metabolic Pathway Metabolism Metalloproteins Metals Microscopy Modification Molecular Molecular Biology Multiprotein Complexes Nature Pathway interactions Physiological Physiology Process Productivity Prokaryotic Cells Proteins Reaction Respiration Role Salmonella enterica Source Spectrum Analysis Sterol O-Acyltransferase Structure Substrate Specificity Techniques Training Vitamin B 12 Vitamins Work antimicrobial biophysical analysis cobamamide fitness human pathogen improved in vivo insight interdisciplinary approach interest microorganism pathogen protein protein interaction small molecule structural biology three dimensional structure

项目摘要

Project Summary/Abstract Understanding the physiological context of the metabolic needs of a pathogen provides a framework for targeted development of antimicrobials. In the case of Salmonella enterica, respiration of ethanolamine in the intestinal environment provides a fitness advantage over commensal microorganisms. Ethanolamine catabolism is a coenzyme B12-dependent pathway that occurs inside a proteinaceous compartment called the ethanolamine (Eut) metabolosome. Ethanolamine is deaminated by ethanolamine ammonia lyase (EAL) to form acetaldehyde, which eventually enters central metabolism as acetyl-CoA. EutA reactivates inactive EAL by removing dysfunctional coenzyme B12 at the expense of ATP. These reactions are needed to trigger the initial step of ethanolamine catabolism hence understanding the interactions required for these processes in the context of the Eut metabolosome would provide opportunities for targeted disruption of the metabolic pathway. There are several gaps of knowledge that need to be filled in so we can improve our understanding f ethanolamine catabolism in this human pathogen. First, the mechanism of catalysis of the adenosyltransferase EutT enzyme that converts vitamin B12 to coenzyme B12 is unknown, thus will be investigated. While the S. enterica EutT is a metalloprotein, EutT homologues in some other pathogens (e.g., Listeria monocytogenes, Clostridum tetani) function without a metal. Metal containing and metal-less Eut enzyme will be studied, and their mechanisms of catalysis compared to elucidate the role of the metal center. Second, it is not understood how coenzyme B12 is delivered to EAL from EutT. Preliminary evidence strongly suggests that EutA mediates the delivery, and that EutT, EutA and EAL may form a complex. A multidisciplinary approach (crystallography, spectroscopy, molecular biology, biochemistry, in vivo genetics, and physiology) will be used to address this complex problem. Collectively, this work will advance our understanding of how cells synthesize and deliver essential coenzymes to the enzymes that use them, in this case inside a cellular compartment.

项目摘要/摘要了解病原体代谢需求的生理背景提供了一个框架有针对性地开发抗微生物药物。在肠道沙门氏菌的情况下，乙醇胺的呼吸与共生微生物相比，肠道环境提供了健康优势。乙醇胺分解代谢是一种依赖辅酶B12的途径，发生在称为乙醇胺(EUT)代谢体。乙醇胺经乙醇胺解氨酶(EAL)脱氨形成乙醛，最终以乙酰辅酶A的形式进入中心代谢。EutA重新激活非活动的EAL 通过以ATP为代价去除功能失调的辅酶B12。这些反应是触发乙醇胺分解代谢的初始步骤，因此理解这些过程所需的相互作用 EUT代谢体的背景将提供有针对性地破坏代谢的机会路径。有几个知识的空白需要填补，这样我们才能提高我们对乙醇胺在这种人类病原体中的分解代谢。第一，腺苷转移酶的催化机制将维生素B12转化为辅酶B12的EutT酶尚不清楚，因此将进行研究。而S。 EutT是一种金属蛋白，EutT在一些其他病原体(如单核细胞增多性李斯特菌、破伤风杆菌)在没有金属的情况下起作用。将研究含金属和无金属的EUT酶，以及通过对其催化机理的比较，阐明了金属中心的作用。第二，不理解辅酶B12如何从EutT传递到EAL。初步证据有力地表明，EutA在交付，以及EutT、EutA和EAL可以形成复合体。多学科方法(结晶学、光谱学、分子生物学、生物化学、体内遗传学和生理学)将被用来解决这个问题复杂的问题。总的来说，这项工作将促进我们对细胞如何合成和传递的理解必需的辅酶与使用辅酶的酶，在这种情况下是在细胞隔间内。