ACETYLCHOLINE BINDING PROTEIN & ACETYLCHOLINE ESTERASE

乙酰胆碱结合蛋白

• 批准号: 7358062
• 负责人: PALMER William TAYLOR
• 金额: $ 1.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7358062
• 关键词: ACETYLCHOLINE; BINDING; PROTEIN; ESTERASE

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. 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 BSome 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 eosi

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。我们通过合成与蜗牛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特性，但我们想检查它是否组装为圆形五聚体，而不是线性四聚体到六聚体。网格上蛋白质的负染色可能会解决这个问题。B部分几年前，埃利斯曼博士和我的实验室采用选择性免疫金标记来定位突触中的乙酰胆碱酯酶。多年来，定位工具已经有了很大的改进，我们现在有了一种小肽，束蛋白，它可以在低 pM 浓度下与乙酰胆碱酯酶相互作用。埃利斯曼实验室开发了一种新型曙红标记技术，大大提高了电子致密材料沉积的分辨率。因此，我们希望利用伊红标记的束蛋白以更高分辨率的化学方法定位突触乙酰胆碱酯酶。我们的实验室拥有制作标记 eosi 的材料和技能