CORE--CHARACTERIZATION OF PROTEINS/RECOMBINANT PROTEINS

核心——蛋白质/重组蛋白质的表征

• 批准号: 6577256
• 负责人: WOLFGANG H FISCHER
• 金额: $ 19.74万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6577256
• 关键词: biomedical facility; follistatin; inhibin; mass spectrometry; protein purification; protein sequence; recombinant proteins

Two essential components of the Projects of this Program will be supported by this Core. The first is the characterization of novel or modified peptides and proteins by Edman based chemical sequencing and mass spectrometric techniques. The second is the expression and purification of proteins that are required by the Projects for biological and biochemical investigations. Protein characterization is an essential part of this Program. The Core provides both state-of-the-art instrumentation and highly trained investigators and support personnel to carry out these characterizations. The Core will characterize novel peptides, proteins or post-translational modifications of known proteins being studied by Program investigators and in particular, mediators involved in actin and inhibin signaling. In addition, we will confirm the sequence and post-translational modifications of recombinant proteins that the Projects generate for use in biological and crystallization studies. Several projects require large quantities of proteins such as activin, activin mutants, inhibin, and soluble fragments of receptors. The Core will generate these proteins by expressing the genes in a suitable system (e.g. baculovirus/insect, mammalian, or yeast cells). The proteins will be purified and characterized using appropriate discriminatory analytical techniques.

本计划项目的两个基本组成部分将由该核心提供支持。第一个是利用基于Edman的化学测序和质谱学技术对新的或修饰的多肽和蛋白质进行表征。第二是生物和生化研究项目所需蛋白质的表达和纯化。蛋白质特性是这项计划的重要组成部分。核心提供最先进的仪器以及训练有素的调查人员和支持人员来进行这些表征。该中心将对项目调查人员正在研究的新的多肽、蛋白质或已知蛋白质的翻译后修饰进行表征，特别是涉及肌动蛋白和抑制素信号的介体。此外，我们将确认该项目产生的用于生物和结晶研究的重组蛋白质的序列和翻译后修饰。有几个项目需要大量的蛋白质，如激活素、激活素突变体、抑制素和受体的可溶性片段。核心将通过在适当的系统(例如杆状病毒/昆虫、哺乳动物或酵母细胞)中表达基因来产生这些蛋白质。这些蛋白质将被提纯，并使用适当的歧视性分析技术进行表征。