PEPTIDE SYNTHESIS CORE

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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. This portion of the parent application requests the resources to support the Core Peptide Synthesis Facility for the major applications in this proposal, as well as for qualified investigators throughout Oklahoma. This Core Facility will primarily construct peptides, ranging from four to twenty amino acids in length using standard, Fmoc chemistry. These peptides will initially be constructed on solid phase supports, which can either be left in their 96-well format or cleaved to free peptides into the fluid phase. The principal investigator of this core has over 16 years of experience using this technique and has published extensively in this area. (Please see attached biosketch and core references.) Peptides constructed by this Facility have previously been used in B and T cell epitope mapping of human and monoclonal sera, inhibition of enzymatic reactivities and mapping of enzymatic active sites. The Core Facility has synthesized peptides for at least two of the key investigators in the past application, as well as providing key resources and preliminary data for the initial funding of two of the additional start-up projects and several of the newly funded collaborative Anthrax and Specialized Center of Research Excellence in Systemic Lupus Erythematosus projects. This Core Facility will serve as a vital resource for several of the current projects, as well as serving the Cell Signaling Core Facility. A large portion of Dr. Chang's proposal is dependent upon solid phase peptides that will be built in the Peptide Core. The core will build screening (8mers) and confirmatory (4-12mers) overlapping peptides of memapsin-2 to identify the key binding sites of memapsin-2 antibodies. Cleaved peptides can also be synthesized for further functional characterization and animal immunization experiments. In addition, the project of Dr. Centola will focus on identifying autoimmune disease-specific cytokine and/or gene expression profiles. Once expression differences are determined, peptides can be generated to confirm findings at the protein level. Based upon our extensive previous experience in the characterization of autoimmune disease phenotypes, the Core will assist Dr. Sawahla with peptide reagents to determine autoimmune specificities and characterize autoantibody binding profiles. Peptides produced for the signaling core will also be used in the projects of Drs. Rodgers and Jackson. As time and resources allow, the Core Peptide Facility will also be available to other investigators from the Foundation, the University of Oklahoma Health Sciences Center, Tulsa University, Oklahoma State University, Oklahoma University and Oklahoma Christian University.