CORE FACILITY RESEARCH PEPTIDE SYNTHESIZER

核心设施研究肽合成器

• 批准号: 6291975
• 负责人: Michael R Yeaman
• 金额: $ 13.21万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6291975
• 关键词: biomedical equipment purchase; peptide chemical synthesis; protein structure function; structural biology

Many important advances in cellular and molecular biology are attributable to structure-function analysis of proteins that .govern function in individual cells or populations of cells. Solid phase peptide synthesis (SPPS) of small peptides, or defined structural, functional, or immunogenic domains of larger proteins, has enabled characterization of structure-activity relationships (SAR) within these molecules that could not be achieved otherwise. The increasing reliance on peptide synthesis by an array of investigators at the Harbor-UCLA Research and Education Institute is the basis for the current effort to acquire a combinatorial SPPS instrument, to expand and modernize the capabilities of the Biopolymer Synthesis Core Facility at Harbor-UCLA REI. As an integral component of this Core Facility, the proposed instrument is adaptable to a wide diversity of peptide syntheses. Thus, peptides produced by this instrument will serve as excellent reagents for a broad array of biomedical research projects. For example, such peptides are invaluable for conducting SAR studies, examining peptide- target cell interactions by flow cytometry, and localization of peptides in situ. Likewise, SPPS enables the production of peptides containing unusual alkylated, alpha,beta-dehydro, and D-enantiomer amino acids crucial for studying conformation as it relates to peptide or protein function. Contemporary expression systems and site-directed mutagenesis approaches are not feasible for routine and rapid production of such peptides. The combinatorial synthesis capability of the proposed instrument will enhance the efficiency of our Core Facility by enabling simultaneous synthesis of multiple, distinct peptides. Applications for unlabeled or labeled peptide reagents to be generated by the proposed instrument for use in NIH-sponsored projects include: 1) protein or peptide functional domain scanning; 2) evaluation of key SAR within proteins and peptides using non-radioactive 13 C- and 15-N-peptide libraries and FTIR or NMR spectroscopy; 3) screening of expression libraries or mutant clones probed with labeled or unlabeled peptides; 4) design and synthesis of peptides with antimicrobial, antitumor, or immunopotentiating properties for eventual therapeutic evaluation; and 5) synthesis of antigen or diagnostic peptide reagents to detect or characterize immunogen targets for vaccine development. Operation and management of the instrument will be streamlined using a simple algorithm coordinating the entire process from on-line peptide synthesis request, to delivery of quality-controlled peptides to investigators, to recharge accounting. The Harbor-UCLA REI is maximally supportive of the long-term success of the instrument as part of the Biopolymer Synthesis Core Facility. Therefore, the proposed instrument will significantly enhance the efficiency and diversity of SPPS required for NIH-sponsored and hypothesis-driven research.

细胞和分子生物学的许多重要进展可归因于对控制单个细胞或细胞群功能的蛋白质的结构功能分析。小肽的固相肽合成 (SPPS) 或较大蛋白质的明确结构、功能或免疫原性结构域，使得能够表征这些分子内的结构活性关系 (SAR)，这是其他方法无法实现的。 Harbor-UCLA 研究和教育研究所的一系列研究人员对肽合成的日益依赖是当前努力购买组合 SPPS 仪器的基础，以扩展和现代化 Harbor-UCLA REI 生物聚合物合成核心设施的能力。作为该核心设施的一个组成部分，拟议的仪器适用于各种肽合成。因此，该仪器产生的肽将作为广泛的生物医学研究项目的优秀试剂。例如，此类肽对于进行 SAR 研究、通过流式细胞术检查肽-靶细胞相互作用以及肽原位定位非常有价值。同样，SPPS 能够生产含有不常见的烷基化、α、β-脱氢和 D-对映体氨基酸的肽，这些氨基酸对于研究与肽或蛋白质功能相关的构象至关重要。现代表达系统和定点诱变方法对于此类肽的常规和快速生产是不可行的。拟议仪器的组合合成能力将通过同时合成多种不同的肽来提高我们核心设施的效率。由拟议仪器生成的未标记或标记肽试剂用于 NIH 资助的项目的应用包括： 1) 蛋白质或肽功能域扫描； 2) 使用非放射性 13 C 和 15-N 肽库以及 FTIR 或 NMR 光谱评估蛋白质和肽内的关键 SAR； 3)筛选用标记或未标记肽探测的表达文库或突变克隆； 4) 设计和合成具有抗菌、抗肿瘤或免疫增强特性的肽，用于最终的治疗评估； 5) 合成抗原或诊断肽试剂以检测或表征疫苗开发的免疫原靶点。该仪器的操作和管理将使用一种简单的算法来简化，该算法协调从在线肽合成请求到将质量控制的肽交付给研究人员，再到充值核算的整个过程。 Harbor-UCLA REI 最大限度地支持该仪器作为生物聚合物合成核心设施的一部分的长期成功。因此，拟议的工具将显着提高 NIH 资助的假设驱动研究所需的 SPPS 的效率和多样性。