Identification and characterization of protein interactions represents an informative approach for understanding mechanisms of normal physiological responses in cells of the nervous tissue and ultimately, cellular responses that are associated with disease. A large number of different technologies have been developed over the last decade that have facilitated the identification of relevant protein interactions and allowed characterization of the cellular and molecular biology of these interactions. These methodologies have resulted in the identification of interactions between proteins involved in the migration of neurons along radial glial cells, ligand/receptor interactions during signal transduction, and intracellular interactions between proteins in the secretory pathways of neuronal cells. We propose to organize a Protein Interaction Core that will offer expression of heterologous proteins in both prokaryotic and eukaryotic expression systems, vector design and selection, biochemical and imaging screening of protein interactions, and more advanced methodologies including two-hybrid screening systems. The goal of the Core Laboratory is to accelerate the identification and characterization of biologically important protein interactions without the need to establish these methodologies in individual laboratories and to facilitative collaborative interactions between NINDS supported investigators at UAB with similar research interests. It is anticipated that the Core Laboratory personnel will assist investigators in selection of appropriate screening systems, generate expression vector constructs for screening assays, and provide troubleshooting and technical assistance in the screening procedures. In addition, the Core Laboratory will encourage the exchange of reagents and more importantly, develop an information sharing resource that will permit individual investigators to share relevant experience with protein expression systems and interacting protein screening systems with other NIIDS supported investigators at UAB and elsewhere. Overall, we believe the proposed core laboratory will limit redundancy in development of these methodologies and permit more efficient use of the resources provided by NINDS. Ultimately, we believe these methodologies and services will facilitate the identifications of relevant protein interactions and assignment of protein function in the complex biologic systems that are goals within the research programs of NINDS supported investigators at UAB.
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