Structural characterization of MCE mediated lipid transport across the Mycobacterial cell envelope

MCE 介导的脂质跨分枝杆菌细胞包膜转运的结构表征

基本信息

批准号：
10427285
负责人：
Casey Vieni
金额：
$ 3.08万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10427285
关键词：
Actinobacteria class Adopted Antibiotics Architecture Bacteria Binding Binding Sites Biochemical Biochemistry Biological Assay Biological Models Cause of Death Cell membrane Cell surface Cells Cholesterol Communicable Diseases Complement Complex Cryoelectron Microscopy Crystallization Cytoplasm DNA DNA Binding DNA Sequence Drug Delivery Systems Drug resistance Environment Escherichia coli Gene Cluster Genes Genetic Genetic Transcription Genome Genus Mycobacterium Glycolipids Hydrophobicity Immune response Knock-out Lead Life Cycle Stages Light Lipids Maintenance Mammalian Cell Maps Mediating Membrane Molecular Movement Mutation Mutation Analysis Mycobacterium smegmatis Mycobacterium tuberculosis Mycobacterium tuberculosis complex Mycolic Acid Nutrient Operon Organism Pathogenicity Phenotype Play Protein Family Proteins Regulation Resources Role Specificity Structure Suggestion System Testing Tuberculosis Virulence Virulence Factors Work antimicrobial cell envelope design differential expression improved insight lipid transport mutant mycobacterial new therapeutic target novel therapeutics protein complex rational design resistant strain response structural biology sulfolipids tuberculosis treatment uptake

项目摘要

PROJECT SUMMARY / ABSTRACT: The cell envelope of Mycobacterium tuberculosis (Mtb) serves an important role as protection against common antimicrobials and host cell defenses, but also a barrier through which Mtb must transport host derived nutrients. The MCE protein family (originally implicated in Mammalian Cell Entry) is nearly ubiquitous in double-membraned bacteria, and MCE proteins are now believed to act as transporters that facilitate lipid movement between the inner and outer membranes. While Mtb MCE proteins are critical for virulence, their mechanism in Mtb remains unclear. Due to Mtb’s distinct cell envelope Mtb MCE proteins likely adopt a unique architecture to mediate transport. In addition, throughout the course of Mtb infection MCE transporters are believed to be differentially expressed by MCE transcriptional regulators (MceRs). This suggests that environmental sensing in Mycobacteria in turn alters transcription of the mce operons. Using structural biology, biochemistry, and genetic complementation assays, I will provide the first glimpse into the architecture of Mtb Mce protein systems and elucidate how MceRs interact with their operator sequences at the molecular level to regulate mce operons. To do so I propose the following aims: (Aim 1) structure determination of the Mce4 lipid transport system from Mycobacterium smegmatis and (Aim 2) structural characterization and DNA-binding specificity of MceRs. My work will provide the first glimpse at the architecture of the M. smegmatis Mce4 transporter system, as well as the molecular mechanisms of MceR mediated mce operon regulation, which may shed light on the mechanisms of cell envelope maintenance in Mtb and provide a novel target for therapeutics.

项目摘要 /摘要：结核分枝杆菌（MTB）的细胞包膜起着重要的作用常见的抗微生物和宿主细胞防御，也是MTB必须运输宿主的障碍衍生的营养素。 MCE蛋白质家族（最初与哺乳动物细胞进入）几乎无处不在现在据信双重膜细菌和MCE蛋白充当促进脂质的转运蛋白内膜和外膜之间的运动。虽然MTB MCE蛋白对病毒至关重要，但 MTB中的机制尚不清楚。由于MTB的独特细胞信封MTB MCE蛋白可能会采用独特的建筑以调节运输。此外，在MTB感染过程中，MCE转运蛋白是被认为是由MCE转录调节剂（MCERS）的不同表达的。这表明这一点分枝杆菌的环境敏感性反过来改变了MCE操纵子的转录。使用结构生物学，生物化学和遗传填充大师，我将首先瞥见MTB的建筑 MCE蛋白系统并阐明MCER与分子水平上的操作员序列相互作用调节MCE操纵子。为此，我提出以下目的：（目标1）MCE4脂质的结构确定分枝杆菌的运输系统和（AIM 2）结构表征和DNA结合 MCERS的特异性。我的工作将对M. Smegmatis MCE4的建筑产生第一瞥见转运蛋白系统以及MCER介导的MCE歌剧调节的分子机制，这些机制可能会阐明MTB中细胞信封维护的机制，并为疗法。