X-RAY CRYSTAL STRUCTURE OF CLOSTRIDIUM PERFRINGENS BETA 2 TOXIN

产气荚膜梭菌 Beta 2 毒素的 X 射线晶体结构

• 批准号: 7721313
• 负责人: Neela H. Yennawar
• 金额: $ 4.06万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721313
• 关键词: Acute; Animals; Binding; Cell membrane; Clostridium perfringens; Computer Retrieval of Information on Scientific Projects Database; Condition; Data; Data Set; Disease; Exotoxins; Food Poisoning; Funding; Goals; Grant; Human; Institution; Membrane; Necrosis; Proteins; Research; Research Personnel; Resources; Roentgen Rays; Screening procedure; Source; Structure; Techniques; Toxin; Ulcer; United States National Institutes of Health; Vaccines; Work; beef; beta pleated sheet; cytotoxicity; enteritis; insight; lead ion; man; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Beta 2 toxin is a 28kD exotoxin produced by Clostridium Perfringens. It is implicated in necrotic enteritis in man and animals, a disease that is characterized by a sudden acute onset with lethal hemorrhagic mucosal ulceration. It is associated with beef contamination resulting in food poisoning in humans. It is pore forming in host cell membranes leaking essential ions leading to cytotoxicity. Pore forming toxins typically transform from soluble monomeric proteins to oligomers that form transmembrane channels. The membrane insertion segment could be alpha helical or part of a beta sheet. Recent structural studies provide some insight into the conformational changes associated with such proteins on membrane association. Our goal is to understand the function of beta 2 toxin from a structural perspective and identify key residues that aid in membrane binding, antigenicity and pore formation. Ultimately we would like to develop a dairy vaccine from the insight gained from solving the 3-dimensional structure. We have grown crystals of beta 2 toxin from Clostridium Perfringens using LiSO4 and Peg3350 as precipitant. We have worked out the best cryo conditions with these crystals. Screening at our local X-ray source has shown some diffraction upto 4 Angstroms. We would like to a. Collect a native data set b. Try quick halide soaks and collect MAD data in order to solve the structure using MAD techniques

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 β2毒素是由灌注梭状芽胞杆菌产生的28KD外毒素。它与人类和动物的坏死性肠炎有关，这种疾病的特征是突然发作并具有致命的出血性粘膜溃疡。它与牛肉污染有关，导致人类食物中毒。它是在宿主细胞膜中形成的孔，导致细胞毒性渗漏。形成毒素的孔通常从可溶性单体蛋白转变为形成跨膜通道的低聚物。膜插入段可以是α螺旋或β表的一部分。最近的结构研究提供了对与此类蛋白相关的膜关联上的构象变化的一些见解。我们的目标是从结构的角度了解β2毒素的功能，并确定有助于膜结合，抗原性和孔形成的关键残基。最终，我们希望从解决三维结构中获得的见解中开发一种乳制品疫苗。 我们使用LISO4和PEG3350作为沉淀剂从灌注梭菌中从梭状芽胞杆菌中种植了β2毒素的晶体。我们已经用这些晶体制定了最好的冷冻条件。在我们的本地X射线源进行筛选显示了一些衍射量高达4埃。我们想 一个。收集本地数据集 b。尝试快速卤化物浸泡并收集疯狂的数据，以便使用疯狂技术解决结构