STABLE FOLD SELECTION FOR HETEROLOGOUS PROTEINS

异源蛋白质的稳定折叠选择

• 批准号: 2647767
• 负责人: ROBERT F BALINT
• 金额: $ 10万
• 依托单位: PANORAMA RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2647767
• 关键词: biotechnology; chemical stability; conformation; method development; mutant; peptide library; protein engineering; protein folding; recombinant proteins

DESCRIPTION: (Slightly modified from applicant's abstract) Dr. Balint et al. have devised a novel solution to two related problems in protein engineering. (1) Most cDNA and random peptide expression libraries are full of sequences which do not express stable proteins. (2) Production costs for many proteins o industrial or pharmaceutical importance are prohibitive due to suboptimal expression of the recombinant protein in heterologous hosts. Their method is based on the testable hypothesis that if any single domain in a multi-domain protein fails to achieve its native fold, the entire protein will turnover and fail to accumulate. If confirmed this fact could be exploited to enrich cDNA expression libraries for stable expressors of native folds in heterologous hosts. The same principles could also be used to pre-enrich random peptide libraries (RPL) for stable expressors, and to select stable, functional, high-level expressors from mutagenic libraries of pharmaceutical or industrial proteins which are otherwise poorly expressed in heterologous hosts. If true, many library of protein domains, whether cDNA, random peptide, or mutagenic, could be expressed as a two-domain fusion via a flexible linker with a selectable marker domain, by which only those members of the library which can achieve stable folds in vivo would be selected. In Phase I they will test this concept. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述：（对申请人的摘要稍作修改）Balint 博士 等人。为蛋白质中的两个相关问题设计了一种新的解决方案 工程。 (1) 大多数cDNA和随机肽表达文库是 充满不表达稳定蛋白质的序列。 （二）生产情况 许多具有工业或制药重要性的蛋白质的成本是 由于重组蛋白的表达不理想而被禁止 异源宿主。他们的方法基于可检验的假设 如果多结构域蛋白质中的任何单个结构域未能实现其 自然折叠时，整个蛋白质将翻转并且无法积累。 如果这一事实得到证实，可用于丰富 cDNA 表达 异源宿主中天然折叠稳定表达的文库。 同样的原理也可用于预富集随机肽 用于稳定表达子的文库（RPL），并选择稳定的、功能性的、 来自药物或药物诱变文库的高水平表达子 工业蛋白质，否则在异源中表达不佳 主机。 如果属实，许多蛋白质结构域文库，无论是 cDNA，还是随机的 肽或诱变肽可以通过以下方式表达为两个结构域融合： 具有可选择标记域的灵活接头，通过该接头，只有那些 可以在体内实现稳定折叠的文库成员是 已选择。 在第一阶段，他们将测试这个概念。 拟议的商业应用：不可用