Maturation of Bacterial Cell Wall

描述（由申请人提供）：细菌的存活取决于其细胞壁的健康状况。由于细胞壁不可或缺的性质，其组装的生物合成酶和细胞壁本身是几种现有抗生素的目标。细胞壁组装和成熟的最后步骤由细胞质膜表面上的一组称为青霉素结合蛋白（PBP）的酶进行。顾名思义，这些酶被β-内酰胺抗生素抑制（例如，青霉素），并且它们充当这些抗生素的杀菌活性的经验证的靶标。尽管它们的重要性，PBPs的生物化学的许多方面还没有被研究，目前没有其他已知的有效抑制剂，这些酶。该项目在三个具体目标的范围内解决信息缺乏的问题。具体目标1涉及具有DD-转肽酶活性的PBPs。这些酶执行细胞壁的重要交联反应。这些研究建立在Mobashery实验室早期工作的基础上，通过实验和计算分析阐明了酶反应中的微观步骤。这些知识被应用到基于机制的抑制这些酶的策略。具体目标2将通过实验和计算来解决具有DD-羧肽酶活性的PBP的机理细节。这些酶水解处理细胞壁，由此它们调节由DD-转肽酶进行的交联程度。具体目标3阐述了具有转糖基酶和DD-转肽酶活性的双功能PBPs的催化机制，明确探索了在细胞壁生物合成中相互影响的两个活性位点之间的串扰。 公共卫生相关性：青霉素结合蛋白（PBP）是内酰胺类抗生素（如青霉素）的靶点，参与细胞壁的成熟，这是一组对细菌生存至关重要的关键功能。对这些酶的功能细节进行了研究。

DESCRIPTION (provided by applicant): Survival of the bacterium depends on the health of its cell wall. Because of the indispensable nature of the cell wall, the biosynthetic enzymes of its assembly and the cell wall itself are targets of several of the existing antibiotics. The final steps of cell wall assembly and maturation are performed by a group of enzymes referred to as penicillin- binding proteins (PBPs) on the surface of the cytoplasmic membrane. As the name suggests, these enzymes are inhibited by beta-lactam antibiotics (e.g., penicillins), and they serve as validated targets for the bactericidal activity by these antibiotics. Despite their importance, many aspects of biochemistry of PBPs have not been investigated and there are currently no other known effective inhibitors for these enzymes. This project addresses this paucity of information within the scope of three Specific Aims. Specific Aim 1 deals with PBPs with the DD-transpeptidase activity. These enzymes perform the important cross-linking reaction of the cell wall. The studies build on earlier work from the Mobashery lab in elucidation of the microscopic steps in the enzymic reaction, both by experiments and by computational analyses. The knowledge is applied to a strategy for mechanism-based inhibition of these enzymes. Specific Aim 2 will address the mechanistic details of PBPs with the DD-carboxypeptidase activity by both experiments and computation. There enzymes process the cell wall hydrolytically, whereby they moderate the degree of cross-linking that is performed by DD-transpeptidases. Specific Aim 3 addresses the catalytic mechanisms of bifunctional PBPs that possess the transglycosylase and DD-transpeptidase activities, explicitly exploring the cross-talk between the two active sites that would influence one another in biosynthesis of cell wall. PUBLIC HEALTH RELEVANCE: Penicillin-binding proteins (PBPs), targets of ¿-lactam antibiotics (such as penicillin), are involved in maturation of cell wall, a set of critical functions that are important for the survival of bacteria. The studies of the details of the functions of these enzymes are proposed.