PEPTIDE SYNTHESIS CORE

肽合成核心

• 批准号: 7959377
• 负责人: Joel Marvin Guthridge
• 金额: $ 7.23万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959377
• 关键词: Active Sites; Amino Acids; Animals; Anthrax disease; Antibodies; Area; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Binding; Binding Sites; Centers of Research Excellence; Chemistry; Cleaved cell; Computer Retrieval of Information on Scientific Projects Database; Core Facility; Data; Epitope Mapping; Foundations; Funding; Gene Expression; Grant; Health Sciences; Human; Immunization; Institution; Left; Length; Liquid substance; Maps; Mentors; Molecular Profiling; Oklahoma; Parents; Peptide Synthesis; Peptides; Phase; Principal Investigator; Proteins; Publishing; Qualifying; Reagent; Request for Applications; Research; Research Personnel; Resources; Science; Screening procedure; Serum; Signal Transduction; Solid; Source; Specialized Center; Specificity; Systemic Lupus Erythematosus; T-Lymphocyte Epitopes; Techniques; Time; United States National Institutes of Health; Universities; base; beta-site APP cleaving enzyme 1; cytokine; disease phenotype; experience; research study

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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 母申请的这一部分请求资源来支持本提案中主要应用的核心肽合成设施，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 化学构建长度为 4 到 20 个氨基酸的肽。这些肽最初将构建在固相支持物上，固相支持物可以保留在 96 孔格式中，也可以裂解成游离肽进入液相。该核心的首席研究员拥有超过 16 年的使用该技术的经验，并在该领域发表了大量文章。 （请参阅随附的生物草图和核心参考文献。）该设施构建的肽先前已用于人类和单克隆血清的 B 和 T 细胞表位作图、酶反应性的抑制以及酶活性位点的作图。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，并为另外两个初创项目和几个新资助的合作炭疽和炭疽病项目的初始资金提供了关键资源和初步数据。系统性红斑狼疮项目专业卓越研究中心。 该核心设施将作为当前几个项目的重要资源，并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取决于将在肽核心中构建的固相肽。核心将构建 memapsin-2 的筛选（8mers）和验证（4-12mers）重叠肽，以识别 memapsin-2 抗体的关键结合位点。还可以合成切割的肽以用于进一步的功能表征和动物免疫实验。此外，Centola 博士的项目将侧重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的发现。基于我们之前在表征自身免疫性疾病表型方面的丰富经验，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谱。为信号核心生产的肽也将用于博士的项目。罗杰斯和杰克逊。如果时间和资源允许，核心肽设施也将向基金会、俄克拉荷马大学健康科学中心、塔尔萨大学、俄克拉荷马州立大学、俄克拉荷马大学和俄克拉荷马基督教大学的其他研究人员开放。