Ordering UNC-13 and UNC-18 function at the synapse

在突触处排序 UNC-13 和 UNC-18 功能

• 批准号: 6937909
• 负责人: CHRISTOPHER E HOPKINS
• 金额: $ 4.4万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6937909
• 关键词: Caenorhabditis elegans; exocytosis; helminth genetics; mutant; postdoctoral investigator; protein binding; protein protein interaction; protein structure function; synapses; synaptic vesicles; syntaxin

DESCRIPTION (provided by applicant): UNC-18 and UNC-13 are presynaptic proteins involved in the synaptic vesicle exocytosis steps of docking, priming, and fusion. UNC-13 functions in priming, yet UNC-18 function is obscure. To determine if UNC-18 can function in either docking or fusion, the ordering of UNC-13 and UNC-18 interactions will be determined. Four specific aims will be pursued. 1) To implicate UNC-18 in docking, syntaxin-complexes will be purified from wild type and unc-13 mutant C. elegans. An increased UNC-18 in the unc-13 background will implicate UNC-18 in vesicle docking. 2) To test for a fusion role, the synaptobrevin and SNAP-25 content of a syntaxin/UNC-18 co-complex will be measured. If UNC-18 is present in complexes containing the requisite SNARE proteins, then a fusion role is implicated. 3) To detect UNC-18 binding to open syntaxin, constitutively open syntaxin mutants will be TAP tagged and screened for the presence of UNC-18, SNAP- 25 and synaptobrevin. The presence of UNC-18 will demonstrate UNC-18 interacts with the open conformation of syntaxin. If SNAP-25 and synaptobrevin are also present, then a fusion role for UNC-18 will be further supported.

描述（由申请人提供）：突触前蛋白18和突触前蛋白13参与突触囊泡胞吐作用的对接、引发和融合步骤。 β-13在启动中起作用，而β-18的作用尚不清楚。 为了确定歼-18是否可以在对接或聚变中发挥作用，将确定歼-13和歼-18相互作用的顺序。 将实现四个具体目标。 1)为了在对接中暗示nc-18，将从野生型和unc-13突变体C纯化突触融合蛋白复合物。优雅的 在unc-13背景中增加的p18将暗示p18参与囊泡对接。 2)为了测试融合作用，将测量突触融合蛋白/突触融合蛋白-18共复合物的小突触泡蛋白和SNAP-25含量。 如果α-18存在于含有必需的SNARE蛋白的复合物中，则涉及融合作用。 3)为了检测与开放型突触融合蛋白结合的β-18，组成型开放型突触融合蛋白突变体将被TAP标记并筛选β-18、SNAP- 25和小突触泡蛋白的存在。 β-18的存在将证明β-18与突触融合蛋白的开放构象相互作用。 如果SNAP-25和小突触泡蛋白也存在，那么将进一步支持β-18的融合作用。