CORE: PEPTIDE SYNTHESIS CORE FACILITY

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。父申请的这一部分要求资源支持核心肽合成设施，用于本提案中的主要应用，以及俄克拉何马州的合格研究人员。该核心设备将主要构建肽，长度从4到20个氨基酸不等，使用标准的Fmoc化学。这些肽最初将构建在固相支架上，固相支架可以保持96孔格式，也可以裂解成游离肽进入液相。这个核心的主要研究者有超过16年的使用这种技术的经验，并在这一领域广泛发表。（请参阅附件的生物草图和核心参考资料。）该设施构建的肽已被用于人类和单克隆血清的B细胞和T细胞表位定位、酶反应活性抑制和酶活性位点定位。在过去的申请中，核心设施已经为至少两位关键研究人员合成了肽，并为另外两个启动项目和几个新资助的合作炭疽和系统性红斑狼疮卓越研究专业中心项目的初始资金提供了关键资源和初步数据。该核心设施将作为几个当前项目的重要资源，以及为细胞信号核心设施提供服务。张博士的提案的很大一部分是依赖于固相肽，将建立在肽核心。该核心将构建memapsin-2的筛选（8mers）和验证（4-12mers）重叠肽，以确定memapsin-2抗体的关键结合位点。裂解肽也可用于进一步的功能表征和动物免疫实验。此外，Centola博士的项目将专注于识别自身免疫性疾病特异性细胞因子和/或基因表达谱。一旦确定了表达差异，就可以生成肽来确认蛋白质水平上的发现。基于我们之前在自身免疫性疾病表型表征方面的丰富经验，Core将协助Sawahla博士使用肽试剂确定自身免疫性特异性并表征自身抗体结合谱。为信号核心生产的多肽也将用于博士的项目。罗杰斯和杰克逊。在时间和资源允许的情况下，核心肽设施也将提供给基金会、俄克拉何马大学健康科学中心、塔尔萨大学、俄克拉何马州立大学、俄克拉何马大学和俄克拉何马基督教大学的其他研究人员。

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.