Bacterial N-Acetyltransferases: Resistance to Regulation

细菌 N-乙酰转移酶：对调节的抵抗

基本信息

批准号：
9264963
负责人：
John S Blanchard
金额：
$ 54.79万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-05-01 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9264963
关键词：
Academia Acetyl Coenzyme A Acetylation Acetyltransferase Acylation Affect Affinity Chromatography Aminoglycoside Antibiotics Aminoglycoside resistance Anabolism Antibiotics Arginine Attention Bacterial Infections Basic Science Biological Biology Cell Extracts Cessation of life Chemicals Chemistry Chloride Ion Clinical CoASH Complex Coupled Crude Extracts Cyclic AMP Deacetylase Defense Mechanisms Development Drug Industry Drug-sensitive Enzymes Exhibits Familiarity Fatty Acids Funding Generations Goals Human Immune response Individual Infection Ingestion Knock-out Laboratories Laboratory Finding Ligase Link Lysine Metabolism Modification Mutation Mycobacterium smegmatis Mycobacterium tuberculosis Nickel Organism Oxygen Peptides Phase Phosphoric Monoester Hydrolases Post-Translational Protein Processing Production Proteins Reagent Regulation Reporting Resistance Ribosomal Proteins Role Scientist Sirtuins Sulfhydryl Compounds System Technology Testing Translations Tuberculosis Wit analog analytical method assault base cohort cytokine design desulfurization extensive drug resistance fluorophore human mortality immune function inhibitor/antagonist macrophage member mortality non-histone protein novel novel therapeutics pathogen programs promoter public health relevance rapid growth resistant strain response small molecule success therapeutic target thioether three dimensional structure

项目摘要

DESCRIPTION (provided by applicant): 4.2.1 Description: Project Summary and Relevance Tuberculosis still remains the number one cause of human mortality caused by infection with a bacterial pathogen. Estimates of mortality range from 1.4-2.0 million deaths globally per year. Current treatment paradigms, developed for drug sensitive strains of Mycobacterium tuberculosis, are being severely threatened by the emergence of multi- and extensively-drug-resistant strains. We have identified a cAMP-dependent protein acetyltransferase and sirtuin-like deacetylase in M. tuberculosis and have identified a small number of protein substrates for the protein acetyltransferase. These enzymes, when acetylated, lose their enzymatic activity, and when deacetylated, are again active. This reversible modification is likely to be important in the transition from rapid growth to a dormant, non-replicating phase. We will in Aim 1, continue our efforts to identify the substrate "cohort" for the protein acetyltransferase and identify growt conditions in which knock-out strains of the protein acetyltransferase or sirtuin are affected. Wit the development of strains of M. tuberculosis that can be grown in BSL2 laboratories, we propose in Aim 2 to identify the small molecule substrates of the 20 GNAT's in M. tuberculosis. We have developed reagents for this purpose, and will continue studies initiated in the prior period of support. In Aim 3, we wish to examine the host proteins that are substrates for the M. tuberculosis EIS protein (EIS = enhanced intracellular survival), an acetyltransferase implicated in modulating the host response to bacterial infection and macrophage ingestion. Together these aims will provide a detailed description of the acetyltransferase enzymes, and their substrates, in an organism whose metabolism is poorly understood.

描述（由申请人提供）：4.2.1描述：项目摘要和相关性结核仍然是由细菌病原体感染引起的人类死亡率的第一名。死亡率的估计范围为每年全球1.4-20万人死亡。目前的治疗范例是为结核分枝杆菌的药物敏感菌株开发而成的，受到多种和广泛抗药性菌株的出现的严重威胁。我们已经鉴定出依赖性cAMP蛋白乙酰转移酶和类似Sirtuin的脱乙酰基酶在结核分枝杆菌中，并确定了蛋白质乙酰基转移酶的少数蛋白底物。当乙酰化时，这些酶会失去其酶活性，而脱乙酰化则再次活跃。这种可逆的修饰可能在从快速生长到休眠，非复制阶段的过渡中很重要。我们将在AIM 1中继续努力确定蛋白质乙酰转移酶的底物“队列”，并确定蛋白质乙酰基转移酶或SIRTUIN的敲除菌株受到影响。在BSL2实验室中可以生长的结核分枝杆菌菌株的发展，我们在AIM 2中提出，以鉴定结核分枝杆菌中20个GNAT的小分子底物。我们已经为此目的开发了试剂，并将继续在上一期支持时开始的研究。在AIM 3中，我们希望检查宿主蛋白，这些宿主蛋白是结核分枝杆菌EIS蛋白的底物（EIS =增强的细胞内生存），这是一种乙酰基转移酶，涉及调节宿主对细菌感染和巨噬细胞摄入的宿主反应。这些目标将共同提供对乙酰转移酶及其底物的详细描述，该酶在一种新陈代谢的生物体中尚不清楚。