PROTEINS & PEPTIDES ASSOCIATED W/ NEURONAL DENSE CORE VESICLES

蛋白质

• 批准号: 7369209
• 负责人: RICHARD FINE
• 金额: $ 0.05万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7369209
• 关键词: PROTEINS; PEPTIDES; ASSOCIATED; NEURONAL; DENSE

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. Synaptic transmission is facilitated by vesicular trafficking of neurotransmitters. The properties and components of synaptic vesicles have been increasingly studied. We are interested in two types of synaptic vesicles (SV), classic synaptic vesicles (CSV) and dense core vesicles (DCV). Studies from our laboratory and others have shown specific protein and neurochemical co-localization with both CSVs and DCVs relevant to neurophysiology and neuropathophysiology. Our studies utilize the tools of mass spectrometry and cell biology for a detailed proteomic study on the components of SVs using existing methodology such as capillary HPLC, ion exchange chromatography, tandem mass spectrometry, and immunohistochemistry. Early results using tandem mass spectral data of in-gel proteolytically digested proteins from isolated vesicles have confirmed previous results identifying the association of specific ion transporters with SVs. Further results indicate specific kinase association with SVs with measurable activity and, using immunoprecipitation and mass spectrometric methods, we are investigating endogenous kinase substrates. 2D chromatographic techniques (e.g., size exclusion followed by reversed-phase) have also been employed. Many expected proteins (e.g., synapsins, synuclein, syntaxin, and others) and peptides (e.g., tachykinins) have been identified. We also have obtained information and structural analysis on post-translational modifications, including glycosylation and acylation, on proteins and peptides from SVs and DCVs. Selective precipitation of proteins has allowed for detection of small proteins and peptides in much greater detail and has increased the number of neuropeptide assignments. Finally, we are investigating the gangliosides associated with CSVs and DCVs. A detailed study should provide not only valuable proteomic and biochemical information but also foster new method development in the examination of the types of molecules associated with vesicles.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。突触传递是由神经递质的囊泡运输促进的。突触囊泡的性质和组成已越来越多地被研究。我们感兴趣的两种类型的突触囊泡（SV），经典的突触囊泡（CSV）和致密核心囊泡（DCV）。我们实验室和其他实验室的研究表明，与神经生理学和神经病理生理学相关的CSV和DCV均存在特异性蛋白质和神经化学共定位。我们的研究利用质谱和细胞生物学的工具，使用现有的方法，如毛细管HPLC，离子交换色谱，串联质谱和免疫组织化学的SV的组件进行详细的蛋白质组学研究。早期的结果使用串联质谱数据的凝胶蛋白水解消化的蛋白质从分离的囊泡已经证实了以前的结果，确定特定的离子转运蛋白与SV的关联。进一步的结果表明，特定的激酶与SV可测量的活动，并使用免疫沉淀和质谱方法，我们正在调查内源性激酶底物。2D色谱技术（例如，尺寸排阻，然后反相）。许多预期的蛋白质（例如，突触蛋白、突触核蛋白、突触融合蛋白等）和肽（例如，速激肽）。我们还获得了关于SV和DCV蛋白质和肽的翻译后修饰（包括糖基化和酰化）的信息和结构分析。蛋白质的选择性沉淀允许更详细地检测小蛋白质和肽，并增加了神经肽分配的数量。最后，我们正在研究与CSV和DCV相关的神经节苷脂。详细的研究不仅提供有价值的蛋白质组学和生物化学信息，而且还促进了与囊泡相关的分子类型的检查新方法的开发。