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归因于对单个细胞或细胞群中的蛋白质的结构 - 功能分析。较大蛋白质的小肽的固相肽合成（SPP），或确定的结构，功能或免疫原性，使这些分子内的结构活性关系（SAR）的表征无法实现。港口 - UCLA研究与教育研究所对肽合成的肽合成的依赖越来越依赖于当前努力获取组合SPPS仪器的基础，以扩大和现代化生物聚合物合成核心核心设施的能力REI。作为该核心设施不可或缺的组成部分，该提出的仪器适合多种肽合成。因此，该仪器生产的肽将作为一系列生物医学研究项目的出色试剂。例如，这种肽对于进行SAR研究，通过流式细胞仪检查肽 - 靶细胞相互作用以及原位肽的定位是无价的。同样，SPP可以产生含有异常烷基化的肽，α，β-脱氢和D-替代异构体氨基酸，这对于研究构象至关重要，因为它与肽或蛋白质功能有关。当代表达系统和位置定向的诱变方法对于这种肽的常规和快速产生不可行。所提出的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的核心设施的效率。拟议的仪器用于NIH赞助的项目中，用于生成未标记或标记的肽试剂的应用包括：1）蛋白质或肽功能域扫描； 2）使用非放射性活性13 C-和15-N肽库以及FTIR或NMR光谱评估蛋白质和肽内的钥匙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）用抗菌，抗肿瘤或免疫构型特性设计和合成用于最终治疗评估的特性； 5）合成抗原或诊断肽试剂以检测或表征疫苗发育的免疫原靶标。将使用简单的算法来简化该仪器的操作和管理，从而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到调查人员的整个过程，以补充会计。 Harbour-Ucla REI最大程度地支持了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，所提出的仪器将显着提高NIH赞助和假设驱动的研究所需的SPP的效率和多样性。