ANALYSIS OF ACETYLCHOLINE BINDING PROTEIN AND ACETYLCHOLINE ESTERASE

点击翻译按钮获取中文摘要

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.