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：
7759198
负责人：
WILLIAM C WIMLEY
金额：
$ 37.72万
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 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7759198
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）：整体膜蛋白约占所有基因组的25％。那些使用B桶基序的人占革兰氏阴性细菌的4％的基因组，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王国中，在包括病原体的毒力和宿主防御以及许多毒素和毒液的作用中，在包括人类在内的所有王国，包括人类，在包括人类的所有王国中都发现了自我组装成B片的外源蛋白质和肽。例如，金黄色葡萄球菌的炭疽毒素和A-溶血素使用B桶渗透膜。还有许多主要是B片结构的孔形成肽抗生素。在这项正在进行的研究计划中，我们正在更深入地了解合成和生物膜上B片的驱动力和结构功能关系，以设计具有有趣且有用的结构和功能的自组装的B-片肽，以便识别B-Barmbrane Membrane Proteins的结构和功能。我们使用的方法用于设计和工程孔形成的肽以及表征抗菌和细胞毒性活性的方法，具有在抗生素，生物传感器和药物递送领域中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线实现。通过筛选库的库来设计新的孔形成肽的家族，这些库为脂质双层膜中的毛孔组装成毛孔的成员。表征脂质囊泡中孔形成的机理以及肽的结构并交叉表征了孔形成器的抗菌活性。设计和表征孔形成的肽，通过筛选文库的有效，广谱抗菌活性。交叉表征基于囊泡的系统中的抗菌肽，以比较两个系统中的结构和功能的决定因素。使用组合化学和高吞吐量筛选来设计自组装到跨膜蛋白质的B桶孔中的肽。最后，我们将使用基因组学和蛋白质组学来描述革兰氏阴性细菌的B桶蛋白质组织，并验证B桶预测算法。此信息将用于创建潜在外膜蛋白的公共数据库

项目成果

期刊论文数量（30）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Biomolecular engineering by combinatorial design and high-throughput screening: small, soluble peptides that permeabilize membranes.

DOI：
10.1021/ja8017863
发表时间：
2008-07-30
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Rathinakumar, Ramesh;Wimley, William C.
通讯作者：
Wimley, William C.

The prediction and characterization of YshA, an unknown outer-membrane protein from Salmonella typhimurium.

DOI：
10.1016/j.bbamem.2010.09.008
发表时间：
2011-01
期刊：
Biochimica et biophysica acta
影响因子：
0
作者：
Freeman TC Jr;Landry SJ;Wimley WC
通讯作者：
Wimley WC

Release of dengue virus genome induced by a peptide inhibitor.

DOI：
10.1371/journal.pone.0050995
发表时间：
2012
期刊：
PloS one
影响因子：
3.7
作者：
Lok SM;Costin JM;Hrobowski YM;Hoffmann AR;Rowe DK;Kukkaro P;Holdaway H;Chipman P;Fontaine KA;Holbrook MR;Garry RF;Kostyuchenko V;Wimley WC;Isern S;Rossmann MG;Michael SF
通讯作者：
Michael SF

Conformational Fine-Tuning of Pore-Forming Peptide Potency and Selectivity.