Folding and Design of Beta Sheets in Membranes

膜中β片的折叠和设计

基本信息

批准号：
7715396
负责人：
WILLIAM C WIMLEY
金额：
$ 1.64万
依托单位：
TULANE UNIVERSITY OF LOUISIANA
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-04-01 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7715396
关键词：
Algorithms Antibiotics Biological Biological Models Biomedical Engineering Biosensor Biotechnology Chlamydia Chloroplasts Databases Drug Delivery Systems Drug Design Engineering Family Figs - dietary Genome Genomics Genus Mycobacterium Goals Gram-Negative Bacteria Hemolysin Host Defense Human Integral Membrane Protein Libraries Life Lipid Bilayers Lipids Membrane Membrane Proteins Methods Mitochondria Peptide Antibiotics Peptides Proteins Proteome Proteomics Research Screening procedure Staphylococcus aureus Structure Structure-Activity Relationship System Thermodynamics Toxin Venoms Vesicle Virulence anthrax toxin antimicrobial antimicrobial peptide base beta pleated sheet combinatorial chemistry cytotoxic design driving force engineering design high throughput screening interest member pathogen peptide structure programs self assembly

项目摘要

DESCRIPTION (provided by applicant): Integral membrane proteins comprise about 25% of all genomes. Those that use the b-barrel motif comprise up to 4% of the genomes of Gram-negative bacteria, and are also present in mycobacteria, chlamydiae, mitochondria and chloroplasts. Exogenous proteins and peptides that self-assemble into b-sheets in membranes are also found in all kingdoms of life, including humans, where they are involved in pathogen virulence and host defense and in the action of many toxins and venoms. For example, the Anthrax toxin and the a-hemolysin of Staphylococcus aureus use b-barrels to permeabilize membranes. There are also many pore-forming peptide antibiotics that are predominantly b-sheet structure. In this ongoing research program we are pursuing a deeper understanding of the driving forces and structure-function relationships for the self-assembly of b-sheets in synthetic and biological membranes in order to design self-assembling b-sheet peptides that have interesting and useful structures and functions in membranes, and in order to identify b-barrel membrane proteins, predict their structure and engineer their function. The methods we are using for designing and engineering pore-forming peptides and for characterizing antimicrobial and cytotoxic activity have potentially important biotechnology applications in the fields of antibiotics, biosensors and drug delivery. These broad goals will be accomplished through the following lines of experimentation. Design new families of pore-forming peptides by screening libraries for members that self-assemble into pores in lipid bilayer membranes. Characterize the mechanism of pore formation in lipid vesicles and the structure of the peptide and cross characterize the antimicrobial activity of pore formers. Design and characterize pore-forming peptides by screening libraries for potent, broad-spectrum antimicrobial activity. Cross characterize antimicrobial peptides in vesicle-based systems to compare the determinants of structure and function in the two systems. Use combinatorial chemistry and high throughput screening to design peptides that self-assemble into membrane-spanning, protein-like b-barrel pores. Finally we will use genomics and proteomics to delineate the b-barrel proteomes of Gram-negative bacteria and validate b-barrel prediction algorithms. This information will be used to create a public database of potential outer membrane proteins

描述（由申请人提供）：整合膜蛋白约占所有基因组的 25%。使用 b 桶基序的基因占革兰氏阴性细菌基因组的高达 4%，并且也存在于分枝杆菌、衣原体、线粒体和叶绿体中。在包括人类在内的所有生命王国中也发现了在膜中自组装成 b-折叠的外源蛋白质和肽，它们参与病原体毒力和宿主防御以及许多毒素和毒液的作用。例如，炭疽毒素和金黄色葡萄球菌的a-溶血素使用b-桶来透化膜。还有许多成孔肽抗生素主要是b-片层结构。在这个正在进行的研究项目中，我们正在寻求更深入地了解合成膜和生物膜中 B-片层自组装的驱动力和结构功能关系，以便设计具有有趣且有用的自组装 B-片层肽膜中的结构和功能，并为了识别 b 桶膜蛋白、预测其结构并设计其功能。我们用于设计和改造成孔肽以及表征抗菌和细胞毒活性的方法在抗生素、生物传感器和药物输送领域具有潜在的重要生物技术应用。这些广泛的目标将通过以下实验来实现。通过筛选库中自组装到脂质双层膜孔中的成员，设计新的成孔肽家族。表征脂质囊泡中孔形成的机制和肽的结构，并交叉表征孔形成剂的抗菌活性。通过筛选具有有效、广谱抗菌活性的文库来设计和表征成孔肽。交叉表征基于囊泡的系统中的抗菌肽，以比较两个系统中结构和功能的决定因素。使用组合化学和高通量筛选来设计可自组装成跨膜、类似蛋白质 b 桶孔的肽。最后，我们将使用基因组学和蛋白质组学来描绘革兰氏阴性细菌的 b 桶蛋白质组并验证 b 桶预测算法。该信息将用于创建潜在外膜蛋白的公共数据库