Interaction of Botulinum neurotoxins with presynaptic receptor complexes

肉毒杆菌神经毒素与突触前受体复合物的相互作用

• 批准号: 7934978
• 负责人: MICHAEL R BALDWIN
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7934978
• 关键词: Address; Affinity; Affinity Chromatography; Antibodies; Axilla; Binding; Binding Sites; Blepharospasm; Bontoxilysin; Botulism; CCRL2 gene; Categories; Cells; Cervical Dystonia; Clinical; Complement; Complex; Development; Disease; Exhibits; Exocytosis; Family; Gangliosides; Glycosphingolipids; Goals; Human; Hyperhidrosis disorder; Immune Sera; Immunoprecipitation; Integral Membrane Protein; Intervention; Intoxication; Laboratories; Mass Spectrum Analysis; Mediating; Membrane; Molecular; Molecular Models; Motor Neurons; Mutagenesis; Neuromuscular Diseases; Neuromuscular Junction; Neurons; Neurotransmitters; One-Step dentin bonding system; Paralysed; Physiological; Presynaptic Receptors; Process; Protein Binding; Protein Isoforms; Proteins; Proteomics; Protocols documentation; Recombinants; Role; SNAP receptor; Serotyping; Site; Site-Directed Mutagenesis; Skeletal Muscle; Structure; Surface; Synaptic Vesicles; Techniques; Testing; Therapeutic; Toxin; Toxoids; Tropism; Vesicle; base; biodefense; improved; insight; member; molecular modeling; novel; novel therapeutics; presynaptic; protein complex; receptor; receptor binding; synaptotagmin; tetanospasmin; tool

Botulism is a neuroparalytic disease that can weaken or paralyze skeletal muscle. The disease is caused by intoxication with one of seven serotypes of botulinum neurotoxin (types A - G). Botulinum neurotoxins (BoNTs) are the most toxic protein toxins of humans and are classified as category A select agents. BoNTs intoxicate neuromuscular junctions through a multistep process involving (a) neuronal cell-binding, (b) internalization into acidic compartments, (c) membrane translocation from acidic compartments, and (d) target recognition and catalytic cleavage of neuronal SNARE proteins required for synaptic vesicle exocytosis. BoNTs are thought to bind to the surface of neurons via a dual receptor mechanism in which the physiologic receptor is a complex composed of gangliosides and protein(s) dual receptors. Using a recombinant receptor binding domain of BoNT, a one-step isolation protocol showed that the BoNT neuronal receptor is a component of a presynaptic receptor complex. The aims of this study will: Aim 1, identify the protein components of the presynaptic BoNT receptor complex. This will be achieved by proteomics and mass spectrometry techniques complemented by immunoprecipitation approaches using BoNT-specific and receptor-specific antibodies. Mm 2 will study the interaction tietween BoNTs and the presynaptic BoNT receptor complex. Utilizing the crystal structures of BoNT receptor binding domains (HCRs), targeted mutagenesis of the HCR domain will be employed to identify the toxin- neuronal receptor interaction sites. Identification of the neuronal receptors for the different serotypes of BoNTs will provide insight and opportunities for the development of novel therapies to inhibit against BoNTs intoxication. Similarly, receptor identification will expand the impact of these studies beyond biodefense, by contributing to improved clinical therapeutic protocols that utilize the BoNTs.

肉毒中毒是一种神经麻痹性疾病，可以削弱或瘫痪骨骼肌。这种疾病是由 肉毒杆菌神经毒素的七种血清型之一（A - G型）中毒。肉毒杆菌神经毒素 （BoNT）是人类毒性最大的蛋白质毒素，被分类为A类选择剂。BoNTs 通过涉及（a）神经元细胞结合，（B） 内化到酸性区室中，（c）从酸性区室的膜易位，和（d）突触囊泡胞吐所需的神经元SNARE蛋白的靶向识别和催化裂解。BoNT被认为通过双受体机制结合到神经元表面，其中生理受体是由神经节苷脂和蛋白质双受体组成的复合物。使用重组受体结合结构域的BoNT，一步分离协议表明，BoNT神经元受体是一个组件的突触前受体复合物。本研究的主要目的是：目的1、鉴定突触前BoNT受体复合物的蛋白组分。这将通过蛋白质组学和质谱技术，辅以使用BoNT特异性和受体特异性抗体的免疫沉淀方法来实现。MM 2将研究BoNT与突触前BoNT受体复合物之间的相互作用.利用BoNT受体结合结构域（HCR）的晶体结构，HCR结构域的靶向诱变将用于鉴定毒素-神经元受体相互作用位点。 不同血清型BoNT的神经元受体的鉴定将提供见解和 开发新疗法以抑制BoNT中毒的机会。同样，受体鉴定将扩大这些研究的影响，超越生物防御，通过促进改善利用BoNT的临床治疗方案。