CORE: PEPTIDE SYNTHESIS CORE FACILITY

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。父应用程序的这一部分请求资源，以支持本提案中的主要应用程序的核心肽合成设施，以及整个俄克拉何马州的合格调查人员。这个核心设施将主要使用标准的FMOC化学来构建长度从4到20个氨基酸的多肽。这些多肽最初将构建在固相载体上，这些固相载体既可以保持96孔格式，也可以裂解成游离肽进入流体相。这一核心的首席研究员有16年以上使用这项技术的经验，并在这一领域发表了大量著作。(请参阅所附的生物草图和核心参考资料。)由该设备构建的多肽曾被用于绘制人和克隆血清的B和T细胞表位图，抑制酶反应活性和绘制酶活性部位图。核心基金在过去的申请中为至少两名关键研究人员合成了多肽，并为另外两个启动项目和几个新资助的系统性红斑狼疮炭疽和专门研究卓越研究中心项目的初始资金提供了关键资源和初步数据。这一核心设施将作为当前几个项目的重要资源，并为小区信令核心设施提供服务。张博士的建议很大程度上依赖于将构建在多肽核心中的固相多肽。核心将建立筛选(8mer)和确证(4-12mer)的Memapsin-2重叠多肽，以确定Memapsin-2抗体的关键结合部位。还可以合成裂解多肽，用于进一步的功能鉴定和动物免疫实验。此外，Centola博士的项目将专注于识别自身免疫性疾病特有的细胞因子和/或基因表达谱。一旦确定了表达差异，就可以产生多肽来在蛋白质水平上证实研究结果。基于我们以往在确定自身免疫性疾病表型方面的丰富经验，Core将协助Sawahla博士使用多肽试剂来确定自身免疫的特异性并确定自身抗体结合的特征。为信号核心生产的多肽也将用于罗杰斯博士和杰克逊博士的项目。在时间和资源允许的情况下，核心肽设施也将提供给来自基金会、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.