Essential Autolysin Activity in Gram-positive Bacteria

革兰氏阳性细菌中的基本自溶素活性

• 批准号: 9035406
• 负责人: HENDRIK SZURMANT
• 金额: $ 12.08万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9035406
• 关键词: Anabolism; Animal Model; Antibiotics; Autolysin; Bacillus subtilis; Biochemical; Cell Wall; Cells; Complement; Complex; Cytoplasm; Cytoplasmic Protein; Databases; Enzymes; Feedback; Fluorescence; Genetic; Goals; Gram-Positive Bacteria; Health; Holoenzymes; Homeostasis; Human; Immune system; Immunofluorescence Immunologic; Knowledge; Learning; Maintenance; Membrane; Modeling; Molecular; Molecular Analysis; Molecular Genetics; Multi-Drug Resistance; Nature; Pathway interactions; Peptidoglycan; Physiological; Post-Transcriptional Regulation; Process; Proteins; Publications; Publishing; Regulation; Regulatory Pathway; Research; Role; Signal Transduction; Skeleton; Structure; System; Techniques; Technology; Work; antimicrobial; base; cell growth; genetic regulatory protein; innovation; insight; macromolecule; multi-drug resistant pathogen; new therapeutic target; optimism; programs; protein complex; research study; structural biology; success; yeast two hybrid system

DESCRIPTION (provided by applicant): The bacterial cell wall is a formidable macromolecule that circumferences the cell. In the past decade a general understanding has been gained for model organisms on the proteins, enzymes and their activities that are involved in the biosynthesis and the restructuring of the cell wall. A main conclusion derived from these studies is that the process of cell wall maintenance is extremely complex and requires the cumulative interactions of many of the involved proteins. It has been suggested that the cell wall biosynthesis machinery exists in the form of a supermolecular holoenzyme, a conclusion that is based on the results from numerous fluorescence localization studies that discovered an interdependence of subcellular localization for these enzymes. Despite these many advances much remains to be learned about the transcriptional feedback loops and posttranscriptional regulations of the involved proteins and the physiologic importance for survival. Our work on the essential WalRK signal transduction system of the Gram-positive bacteria has identified this regulatory system to connect cellular growth with the ensuing necessity for cell wall restructuring. As a consequence of these studies essential autolysin activities have been unveiled that are subject to transcriptional and posttranscriptional control and that might be targeted by the innate immune system. Building on this knowledge we propose here to discover the concerted interplay between the many players that are involved in Bacillus subtilis cell wall homeostasis as a model for Gram-positive cell wall maintenance. The identification of molecular and structural details of the protein interactions involved in cell wall restructuring is hampered y the transient nature of these interactions and by the fact that the involved proteins span all thre compartments of the cell from cytoplasm to membrane to extra-cytoplasmic space. To identify interaction parameters between interacting proteins we recently developed a computational approach. The approach relies on extensive and ever growing sequence databases, to elucidate contact residue information between interacting proteins. Recent publications and preliminary results strongly support the notion that this technology is generally applicable to decipher protein interaction information, and will now be applied to the study of important transient interaction that govern cell wall maintenance and restructuring.

描述（由申请人提供）：细菌细胞壁是绕细胞绕过细胞的强大的大分子。在过去的十年中，人们对蛋白质，酶及其活性的模型生物有了一般的了解，这些酶及其活性涉及生物合成和细胞壁的重组。这些研究得出的一个主要结论是，细胞壁维持过程非常复杂，需要许多涉及蛋白的累积相互作用。已经提出，细胞壁生物合成机制以超分子全酶的形式存在，这一结论基于许多荧光定位研究的结果，这些研究发现了这些酶的亚细胞定位的相互依存关系。尽管有许多进展，还有很多尚待了解有关蛋白质的转录反馈回路和转录后法规以及生存的生理重要性。我们在革兰氏阳性细菌的基本Walrk信号转导系统上的工作已经确定了该调节系统，以将细胞生长与随之而来的细胞壁重组的必要性联系起来。由于这些研究的结果，基本的自动蛋白素活动已揭幕，这些活动受转录和转录后控制的影响，并且可能是先天免疫系统的目标。在此知识的基础上，我们在这里建议发现涉及枯草芽孢杆菌细胞壁稳态的许多玩家之间的一致相互作用，作为革兰氏阳性细胞壁维护的模型。在细胞壁重组中涉及的蛋白质相互作用的分子和结构细节的鉴定受到了这些相互作用的瞬时性质，并且涉及的蛋白质跨越了从细胞质量到膜到膜外质质空间的所有细胞的所有隔室。为了确定相互作用蛋白质之间的相互作用参数，我们最近开发了一种计算方法。该方法依赖于广泛而不断增长的序列数据库，以阐明相互作用蛋白之间的接触残留信息。最近的出版物和初步结果强烈支持以下观点：该技术通常适用于破译蛋白质相互作用信息，现在将应用于研究细胞壁维持和重组的重要瞬态相互作用的研究。