Supramolecular Complexes That Mediate Pneumococcal PG Biosynthesis and Virulence

介导肺炎球菌 PG 生物合成和毒力的超分子复合物

基本信息

批准号：
8507826
负责人：
MALCOLM E. WINKLER
金额：
$ 38.22万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8507826
关键词：
ATP Hydrolysis ATP-Binding Cassette Transporters Accounting Address Anabolism Antibiotics Binding Biochemical Carboxypeptidase Cell Shape Cell Size Cell Wall Cell division Cell surface Cellular biology Clinical Complex Coupled Couples Data Enzymes Genetic Goals Gram-Positive Bacteria Human Hydrolysis Immune response Infection Knowledge Lactams Link Location Mediating Membrane Proteins Microscopic Modeling Multi-Drug Resistance N-Acetylmuramoyl-L-alanine Amidase Paper Passive Immunity Pathogenesis Penicillin-Binding Proteins Peptides Peptidoglycan Peripheral Physiology Play Pneumococcal Colonization Process Property Proteins Resistance development Role Scaffolding Protein Shapes Staging Streptococcus Streptococcus pneumoniae Structure Teichoic Acids Tertiary Protein Structure Testing Time Vancomycin Virulence Virulence Factors Work base capsule clinically relevant combat expectation feeding genetic regulatory protein innovation killings meetings mutant novel vaccines pathogen protein complex respiratory scaffold sortase vaccine candidate vaccine development

项目摘要

DESCRIPTION (provided by applicant): Streptococcus pneumoniae (pneumococcus) is an extremely serious human respiratory pathogen that kills well over two million people annually worldwide. Multidrug resistance is increasing in S. pneumoniae clinical isolates at an alarming rate. Many clinically relevant antibiotics, including ¿-lactams and vancomycin, target peptidoglycan (PG) biosynthesis. PG forms the major rigid structure in the cell wall that determines cell shape and size and serves as the scaffolding onto which other pneumococcal virulence factors are covalently attached, including capsule, teichoic acids, and sortase-transferred proteins. Despite its importance to physiology and pathogenesis, little is known about the supramolecular protein complexes that mediate PG biosynthesis on the cell surface of S. pneumoniae and other ellipsoid-shaped ovococcus Gram-positive pathogens. The long-term goal of this project is to fill in this major knowledge gap about the locations, interactions, regulatory dynamics, and functions of the supramolecular protein complexes that mediate pneumococcal PG biosynthesis. This proposal is based on a large body of new papers and unpublished data that demonstrates numerous unique and unexpected properties of PG biosynthesis in S. pneumoniae. PG biosynthesis is a broad topic that encompasses both PG synthesis and PG remodeling by hydrolysis. This five- year proposal consists of four synergistic Specific Aims that address some of the most important outstanding problems in ovococcus PG biosynthesis. These four Aims are conceptually linked to the central hypothesis that PG synthesis and PG remodeling enzymes function as dynamic supramolecular complexes, whose activities and interactions are choreographed with each other and with cell division. These Aims were chosen, because they will yield fundamental principles about PG biosynthesis, are supported by strong new data, are experimentally tractable, feed into each other, and will have high impact on the field. Related Aims 1 and 2 will elucidate how penicillin binding proteins (PBPs) are localized, activated, and tied to stages of cell division through interactions with a small number of essential master organizer proteins. Aim 3 will test the hypothesis that PG hydrolysis involved in PG remodeling is coupled directly to cell division. Aim 4 will determine how PG hydrolases modulate the supply of PG pentapeptide substrates used by PBPs and whether PG peptides play roles in organizing PG synthesis. A comprehensive strategy that combines results from innovative genetic, biochemical, cell biology, and microscopic approaches with those from colonization and infection models will be used to meet these Aims for this primary bacterial pathogen. Results from this proposal will challenge and expand paradigms and models about PG biosynthesis in S. pneumoniae and other ovococcus pathogens. Since the cell surface is critical to pneumococcal virulence and extracytoplasmic proteins are accessible and druggable, there is an expectation that some of the critical PG synthesis and remodeling proteins studied in this proposal will emerge as new antibiotic and vaccine candidates.

描述（由适用提供）：肺炎链球菌（肺炎球菌）是一种非常严重的人类呼吸道病原体，每年在全球范围内杀死超过200万人。肺炎链球菌临床分离株的多药耐药性以惊人的速度增加。许多与临床相关的抗生素，包括 - lactams和万古霉素，靶向肽聚糖（PG）生物合成。 PG在细胞壁中形成了确定细胞形状和大小的主要刚性结构，并用作其他肺炎球菌病毒因子共价附着的脚手架，包括胶囊，Teichoic酸和分类酶转移的蛋白质。 Despite its importance to physiology and pathogenesis, little is known about the supramolecular protein complexes that mediate PG biosynthesis on the cell The long-term goal of this project is to fill in this major knowledge gap about the locations, interactions, regulatory dynamics, and functions of the supramolecular protein complexes that mediate pneumococcal PG biosynthesis.该提案基于大量新论文和未发表的数据，这些数据表明S. PG生物合成中PG生物合成的众多独特和意外特性是一个广泛的主题，它涵盖了PG合成和通过水解进行PG重塑。这项为期五年的提案由四个协同特定目的组成，这些目的解决了Ovococcus PG生物合成中一些最重要的问题。这四个目标在概念上与中心假设相关，即PG合成和PG重塑酶起动态超分子复合物的作用，后者的活动和相互作用彼此编排，并与细胞分裂进行了编排。之所以选择这些目标，是因为它们将产生有关PG生物合成的基本原理，并得到强大的新数据的支持，可以在实验上进行实验，彼此相互进食，并对现场产生很大的影响。相关目标1和2将阐明青霉素结合蛋白（PBP）如何通过与AIM 3的相互作用来局部，激活并与细胞分裂的阶段相关，这将检验以下假设：PG水解参与PG重塑的PG水解直接与细胞分裂偶联。 AIM 4将确定PG水解酶如何调节PBPS使用的PG五肽底物的供应以及PG Petites是否在组织PG合成中起作用。由创新的遗传，生化，细胞生物学和微观方法与殖民化和感染模型的综合策略结合起来，以实现这些主要细菌病原体的这些目标。该建议的结果将挑战和扩大有关肺炎链球菌和其他卵球菌病原体中PG生物合成的范例和模型。由于细胞表面对肺炎球菌病毒至关重要，并且可及性蛋白质可及可及性蛋白质，因此人们期望在此提案中研究的一些关键的PG合成和重塑蛋白质将以新的抗生素和疫苗候选者出现。