Structural and functional studies of botulinum neurotoxin

肉毒杆菌神经毒素的结构和功能研究

• 批准号: 8590196
• 负责人: Rongsheng Jin
• 金额: $ 34.7万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-15 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8590196
• 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

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（由申请人提供）：肉毒杆菌神经毒素（BONTS）是人类已知的最有毒物质之一，小鼠中有50％的致命剂量（LD50）为1 ng/kg。因此，BONT代表了主要的生物恐怖威胁（Arnon等，2001； Gill，1982）。矛盾的是，含有散发的药物和化妆品，例如肉毒杆菌毒素（R），Dysport（R），Xeomin（R），CBTXA（R），肌无菌（R）和神经斑boc（R），已成功地使用了多种神经学，耳鼻喉科学，口感学，尿液学，尿液学，尿液学，尿液学，尿液学，尿液学，尿液学，尿液学，并有效，并有效（Jankovic，2004； Truong和Jost，2006）。 2010年10月，FDA批准了肉毒杆菌毒素（R）治疗慢性偏头痛。 BONT的毒性和治疗功能都取决于进入神经元的通用机制，裂解蛋白质，介导关键神经递质的胞吐作用，然后瘫痪受影响的肌肉。 BONT由肉毒梭状芽胞杆菌作为大祖细胞毒素复合物（PTC）产生，其中包括称为神经毒素相关蛋白（NAPS）（Montecucco和Schiavo，1995）的辅助梭菌蛋白（NAPS）。临床配方也由BONT PTC组成。意外骨中毒主要是通过口服摄入污染的食品而发生的。人们认为午睡可以保护BONT避免胃肠道（GI）的敌对环境，并介导跨上皮细胞屏障的毒素转运，这是中毒的第一步。但是，控制这些过程的基本分子机制知之甚少。该提案的目的是阐明BONT PTC的结构和功能，包括午睡保护BONT的机制，PTC组装下的调节机制以及小睡在调节BONTS和宿主细胞之间相互作用的结构基础。该提案的重点是BONT（BONT/A）的血清型A，因为它是生物恐怖主义的主要关注点，并且是所有BONT血清型中最常用的药物。我们将结合包括X射线晶体学的技术，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。如果成功的话，拟议的锅将指导新型治疗剂的设计，以治疗和/或预防肉毒杆菌。我们的工作还可以为肉毒杆菌毒素（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功效并提出新的临床应用。