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. Progress:
Specific Aim : Develop a soluble amino terminal binding domain of the serotonin type 3 receptor with ligand specificities identical or similar to the 5-HT3R. This project began May 1, 2004 with the following first year goals: 1. Establish the PIs laboratory. 2. Hire 2 graduate students and a research Associate 3. Establish stably expressing cell lines for the Acetylcholine Binding Protein (AChBP). All first year goals have been accomplished. Two graduate students were hired over the past 12 months. One student began working in August 2004 and a second in January 2005. Both students are working on this project. A Research Associate (Dr. Jestina Kusina) was hired in March 2005 and has begun work in the laboratory primarily focusing on methods development. All laboratory equipment has been purchased and is currently being employed in the project. Active research began in November 2004 and has been focused on developing the expression systems and protein purification protocols outlined in the proposal. We have developed stably transfected cells lines expressing the AChBP. The purification and characterization of this protein will begin shortly. AChBP pharmacology will be characterized by 96 well assays and Surface Plasmon Resonance (SPR). Dr. Kusina is currently developing the SPR assays for this purpose. Syntheses of the AChBP/5-HT3R chimeras are currently underway and should be complete by the time we have developed the supporting protocols. Project goals have been expanded slightly to include the use of the expressed AChBP as a biosensor in drug development. This will enable us to make use of the AChBP expression system and SPR to evaluate the pharmacological specificity of this protein. Since the AChBP is being used as a structural template for many other ligand-gated ion channels, these data should constitute a valuable contribution to the LGIC field. We are also exploring the use of an AChBP/SPR sensor for detection of anti-nAChR antibodies in Myasthenia Gravis.
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