ANALYSIS OF ACETYLCHOLINE BINDING PROTEIN AND ACETYLCHOLINE ESTERASE

乙酰胆碱结合蛋白和乙酰胆碱酯酶的分析

• 批准号: 7601095
• 负责人: PALMER William TAYLOR
• 金额: $ 0.43万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601095
• 关键词: Acetylcholine; Acetylcholinesterase; Binding Proteins; Chemistry; Chromosome Pairing; Computer Retrieval of Information on Scientific Projects Database; Deposition; Electrons; Eosine Yellowish; Extracellular Domain; Funding; Grant; Homologous Gene; Institution; Label; Laboratories; Ligands; Localized; Negative Staining; Nicotinic Receptors; Peptides; Property; Proteins; Research; Research Personnel; Resolution; Resources; Snails; Source; Specificity; Structure; Synapses; Techniques; United States National Institutes of Health; esterase; fasciculin; improved; novel; skills; tool

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. Analysis of the structure of an acetylcholine binding protein. We have have prepared the extracellular domain of the nicotinic acetylcholine receptor by synthesizing a gene homologous with the snail binding protein. It shows the appropriate ligand specificity and hydrodynamic properties, but we would like to check to see whether it assembles as a circular pentamer rather than a linear tetramer through hexamer. Negative staining of the protein on a grid may resolve this issue. Part 2: Some years ago Dr. Ellisman's and my laboratory employed selective immunogold labeling to localize acetylcholinesterase in the synapse. The tools for localization have greatly improved aver the years, and we now have a small peptide, fasciculin, that interacts with acetylcholinesterase at low pM concentrations. The Ellisman laboratory has developed a novel eosin labeling technique that greatly improves the resolution of deposition of electron dense materal. Hence, we hope to employ eosin-labeled fasciculin to localize synaptic acetylcholinesterase with higher resolution chemistry. Our laboratory has the materials and skills to make labeled eosin.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 乙酰胆碱结合蛋白的结构分析。我们已经通过合成与snail结合蛋白同源的基因制备了烟碱乙酰胆碱受体的胞外结构域。 它显示了适当的配体特异性和流体动力学性质，但我们想检查它是否组装成环状五聚体，而不是线性四聚体到六聚体。 在网格上对蛋白质进行负染色可以解决这个问题。 第二部分：几年前，Ellisman博士和我的实验室采用了选择性免疫金标记法来定位突触中的乙酰胆碱酯酶。 这些年来，定位的工具已经有了很大的改进，我们现在有了一种小肽，fasciculin，它在低pM浓度下与乙酰胆碱酯酶相互作用。 Ellisman实验室开发了一种新的曙红标记技术，大大提高了电子致密材料沉积的分辨率。 因此，我们希望采用曙红标记的fasciculin定位突触乙酰胆碱酯酶与更高的分辨率化学。我们实验室有材料和技术来制造标记曙红。