Structural and functional studies of botulinum neurotoxin

肉毒杆菌神经毒素的结构和功能研究

基本信息

批准号：
8401135
负责人：
Rongsheng Jin
金额：
$ 14.71万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-15 至 2013-04-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8401135
关键词：
Accounting Adverse drug effect Affect Affinity Architecture Bacteria Binding Biochemistry Bioinformatics Biophysics Bioterrorism Blood Circulation Bontoxilysin Botox Botulinum Toxin Type A Botulism Cells Cellular biology Chronic Cleaved cell Clinical Clostridium botulinum Complex Cosmetics Development Drug Delivery Systems Drug Formulations Elan brand of botulinum toxin type B Environment Enzymes Epithelial Epithelial Cells Exocytosis FDA approved Food Gastrointestinal Diseases Gastrointestinal tract structure Gills Goals Hemagglutinin Ingestion Intestines Intoxication Lead Lethal Dose 50 Mediating Medicine Migraine Molecular Motor Neurons Mus Muscle Neurologic Neuromuscular Junction Neurons Neurotoxins Neurotransmitters Oral Paralysed Pathogenesis Peptide Hydrolases Poisoning Prevention Process Proteins Resolution Role Serotyping Specificity Staging Structure Techniques Testing Therapeutic Toxicology Toxin Transportation Work X-Ray Crystallography base clinical application clinical efficacy design improved insight man molecular mass novel novel strategies novel therapeutic intervention novel therapeutics premature prevent progenitor receptor binding small molecule success urologic weapons

项目摘要

DESCRIPTION (provided by applicant): Botulinum neurotoxins (BoNTs) are among the most poisonous substances known to man, with a 50% lethal dose (LD50) of 1 ng/kg in mice. BoNTs therefore represent a major bioterrorist threat (Arnon et al., 2001; Gill, 1982). Paradoxically, BoNT-containing medicines and cosmetics, such as Botox(R), Dysport(R), Xeomin(R), CBTXA(R), Myobloc(R) and NeuroBloc(R), have been used with great success to treat a variety of neurological, otolaryngological, ophthalmological, urological, dermatological and gastrointestinal disorders (Jankovic, 2004; Truong and Jost, 2006). In October 2010, FDA approved Botox(R) to treat chronic migraine. Both the toxic and therapeutic functions of BoNTs rely on a common mechanism to enter neurons, cleave proteins that mediate exocytosis of key neurotransmitters, and subsequently paralyze the affected muscles. BoNTs are produced by Clostridium botulinum as large progenitor toxin complexes (PTCs) that include auxiliary clostridial proteins known as neurotoxin-associated proteins (NAPs) (Montecucco and Schiavo, 1995). Clinical formulations are also composed of BoNT PTCs. Accidental BoNT poisoning mainly occurs through oral ingestion of tainted food products. NAPs are thought to protect BoNTs against the hostile environment of the gastrointestinal (GI) tract, and to mediate toxin transport across the epithelial cell barriers, the first step of intoxication. However the underlying molecular mechanisms that control these processes are poorly understood. The goal of this proposal is to elucidate the structure and function of BoNT PTCs, including the mechanism by which NAPs protect BoNTs, the regulatory mechanism underlying PTC assembly, and the structural basis by which NAPs regulate the interaction between BoNTs and host cells. The focus of this proposal is on the serotype A of BoNT (BoNT/A), because it is a major concern for bioterrorism and is the most commonly used medicine among all BoNT serotypes. We will use a combination of techniques, including X-ray crystallography, together with biochemistry, biophysics, cell biology, toxicology and bioinformatics. If successful, the proposed wok will guide the design of novel therapeutics for the treatment and/or prevention of botulism. Our work may also provide new insights into the activity and side effects of drugs such as Botox(R) and Dysport(R), which may help improve their clinical efficacy and suggest novel clinical applications.

描述（申请人提供）：肉毒杆菌神经毒素（BoNT）是人类已知的毒性最强的物质之一，对小鼠的 50% 致死剂量（LD50）为 1 ng/kg。因此，BoNT 代表了主要的生物恐怖威胁（Arnon 等人，2001 年；Gill，1982 年）。矛盾的是，含有 BoNT 的药物和化妆品，如 Botox(R)、Dysport(R)、Xeomin(R)、CBTXA(R)、Myobloc(R) 和 NeuroBloc(R)，已被成功用于治疗各种神经科、耳鼻喉科、眼科、泌尿科、皮肤科和胃肠道疾病（Jankovic，2004；Truong 和 Jost， 2006）。 2010年10月，FDA批准Botox(R)用于治疗慢性偏头痛。 BoNT 的毒性和治疗功能都依赖于一种共同的机制来进入神经元，裂解介导关键神经递质胞吐作用的蛋白质，并随后麻痹受影响的肌肉。 BoNT 由肉毒梭菌产生，作为大型祖毒素复合物 (PTC)，其中包括被称为神经毒素相关蛋白 (NAP) 的辅助梭菌蛋白 (Montecucco 和 Schiavo，1995)。临床制剂也由 BoNT PTC 组成。意外 BoNT 中毒主要是通过口服受污染的食品而发生。 NAP 被认为可以保护 BoNT 免受胃肠道 (GI) 的恶劣环境的影响，并介导毒素穿过上皮细胞屏障的转运，这是中毒的第一步。然而，人们对控制这些过程的潜在分子机制知之甚少。该提案的目标是阐明BoNT PTC的结构和功能，包括NAP保护BoNT的机制、PTC组装的调控机制以及NAP调节BoNT与宿主细胞之间相互作用的结构基础。该提案的重点是 BoNT 血清型 A（BoNT/A），因为它是生物恐怖主义的主要关注点，也是所有 BoNT 血清型中最常用的药物。我们将结合使用 X 射线晶体学等技术以及生物化学、生物物理学、细胞生物学、毒理学和生物信息学。如果成功，所提出的锅将指导治疗和/或预防肉毒杆菌中毒的新型疗法的设计。我们的工作还可能为 Botox(R) 和 Dysport(R) 等药物的活性和副作用提供新的见解，这可能有助于提高其临床疗效并提出新的临床应用。