PEPTIDE SYNTHESIS CORE

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 母申请的这一部分要求提供资源，以支持核心肽合成设施用于本提案中的主要应用，以及整个俄克拉荷马州的合格研究人员。该核心设施将主要使用标准Fmoc化学方法构建长度为4至20个氨基酸的肽。这些肽最初将在固相支持物上构建，所述固相支持物可以以其96孔形式保留或裂解成游离肽进入流体相。该核心的主要研究者拥有超过16年的使用该技术的经验，并在该领域发表了大量文章。（请参见随附的生物插图和核心参考文献。）由该设施构建的肽先前已用于人和单克隆血清的B和T细胞表位作图、酶反应性的抑制和酶活性位点的作图。核心设施在过去的申请中为至少两名关键研究人员合成了肽，并为两个额外的启动项目和几个新资助的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。 该核心设施将作为当前几个项目的重要资源，并为Cell Signaling Core Facility服务。 Chang博士的提议的很大一部分取决于将在肽核心中构建的固相肽。该核心将构建筛选（8聚体）和确证（4- 12聚体）的memapsin-2重叠肽，以鉴定memapsin-2抗体的关键结合位点。也可以合成切割的肽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，就可以产生肽以在蛋白质水平上确认发现。基于我们在自身免疫性疾病表型表征方面的丰富经验，核心将协助Sawahla博士使用肽试剂来确定自身免疫特异性并表征自身抗体结合特征。为信号核心产生的肽也将用于罗杰斯和杰克逊博士的项目。随着时间和资源的允许，核心肽设施也将提供给其他研究人员从基金会，俄克拉荷马州健康科学中心，塔尔萨大学，俄克拉荷马州州立大学，俄克拉荷马州大学和俄克拉荷马州基督教大学。

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.