The MmpL3 interactome

MmpL3 相互作用组

• 批准号: 10382791
• 负责人: Georgiana E. Purdy
• 金额: $ 19.25万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-27 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10382791
• 关键词: Adaptor Signaling Protein; Anabolism; Antibiotic Resistance; Binding; Biochemical Genetics; Biogenesis; Cause of Death; Cell Wall; Cell division; Cell membrane; Communicable Diseases; Complex; Cord Factors; Cytoplasm; Data; Drug Targeting; Esters; Future; Glycoconjugates; Hydrophobicity; In Vitro; Lipid Binding; Lipids; Lipoproteins; Membrane; Membrane Proteins; Modeling; Modification; Mycobacterium tuberculosis; Mycolic Acid; Nodule; Pathogenesis; Peptidoglycan; Periplasmic Proteins; Phenotype; Physiology; Protein Family; Proteins; Publications; Resistance; Role; Site; Transferase; Trehalose; Triglycerides; Tuberculosis; Virulence; Waxes; Work; arabinogalactan; base; cell envelope; crosslink; genetic approach; in vivo; interest; lipid transport; mutant; mycobacterial; mycolate; protein complex; therapeutic target; tuberculosis drugs

ABSTRACT Tuberculosis is one of the leading causes of death due to infectious disease despite the availability of anti- tubercular drugs. The cell envelope of Mycobacterium tuberculosis (Mtb) is notable for the abundance of mycolic acids (MAs), which are essential to mycobacterial viability. The mycobacterial cell envelope is extremely hydrophobic, contributes to virulence and antibiotic resistance. Yet, exactly how glycoconjugates and other species-specific lipids are transported across the inner membrane for cell envelope biosynthesis is incompletely understood. Recent work shows that the Mycobacterial membrane protein Large (MmpL) transporters export lipids synthesized in the mycobacterial cytoplasm for incorporation into the cell envelope. These transporters are therefore important for bacterial viability and virulence. MmpL3, the focus of this proposal, is essential and required for transport of trehalose monomycolate (TMM), the precursor of trehalose dimycolate (TDM) and mycolyl arabinogalactan peptidoglycan (mAGP). The exact mechanism of MmpL3 export remains elusive. RND family proteins typically possess adaptor proteins that assist in substrate transport. As presented in our preliminary data and recent publication, we identified several lipoproteins that interact with MmpL3 and MmpL11. We characterized an Mtb lpqN mutant and demonstrated that LpqN has a lipid binding pocket and interacts with the Ag85 mycolyl transferases. Based on these data, we propose a model where mycobacterial lipoproteins form a complex with MmpL protein to promote lipid secretion and localization. Our hypothesis is that MmpL3 interacts with lipoproteins that facilitate export of TMM and its incorporation into the cell envelope. The proposed study will combine biochemical and genetic approaches to identify proteins that complex with MmpL3 to facilitate substrate transport. .

抽象的 结核病是由于传染病而导致死亡的主要原因之一，尽管有抗 结核药物。结核分枝杆菌（MTB）的细胞包膜很有意义 霉菌酸（MAS），这对于分枝杆菌生存力至关重要。分枝杆菌细胞信封是 极度疏水，有助于毒力和抗生素耐药性。然而，恰好是糖缀合物 和其他特异性脂质在整个内膜上运输以进行细胞包膜生物合成是 不完全理解。 最近的工作表明，分枝杆菌膜蛋白大（MMPL）转运蛋白导出脂质 在分枝杆菌细胞质中合成以掺入细胞包膜中。这些转运蛋白是 因此对于细菌的生存能力和毒力很重要。 MMPL3是本提案的重点，是必不可少的，并且 海藻糖单切酸（TMM），海藻糖二酸（TDM）和 霉菌基因共糖肽肽聚糖（MAGP）。 MMPL3导出的确切机制仍然难以捉摸。 RND家族蛋白通常具有有助于底物运输的衔接蛋白。如所示 我们的初步数据和最新出版物，我们确定了几种与MMPL3相互作用的脂蛋白 MMPL11。我们表征了MTB LPQN突变体，并证明LPQN具有脂质结合袋，并且 与AG85霉菌基转移酶相互作用。基于这些数据，我们提出了一个模型，其中分枝杆菌 脂蛋白与MMPL蛋白形成复合物，以促进脂质分泌和定位。 我们的假设是MMPL3与脂蛋白相互作用，脂蛋白有助于出口TMM及其 掺入细胞包膜中。拟议的研究将结合生化和遗传方法 鉴定与MMPL3复合物的蛋白质以促进底物运输。 。