New chaperones for folding and assembly of proteins

用于蛋白质折叠和组装的新伴侣

• 批准号: 7671192
• 负责人: FRANK Thomson ROBB
• 金额: $ 17.56万
• 依托单位: FIDELITY SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7671192
• 关键词: Archaea; Area; Biochemical Reaction; Biological Assay; Biology; Biotechnology; Cells; Collaborations; Complement; Engineering; Ensure; Environment; Enzymes; Escherichia coli; Event; Exhibits; Fluorescence; Genetic Transcription; Genome; Goals; Green Fluorescent Proteins; Healthcare; Homologous Gene; In Vitro; Inclusion Bodies; Institutes; Isopropyl Thiogalactoside; Life; Maryland; Mediating; Metabolic; Methods; Molecular Chaperones; Molecular Conformation; Monitor; Operon; Pathway interactions; Phase; Physiological; Play; Process; Production; Productivity; Property; Protein Deficiency; Proteins; Proteomics; Protocols documentation; Quality Control; Reagent; Recombinant Proteins; Relative (related person); Reporter; Resistance; Role; Sodium Chloride; Solubility; Stress; System; Technology; Temperature; Testing; Time; Universities; base; chaperonin; chromophore; clinical application; cold temperature; combinatorial; improved; in vivo; innovation; novel; novel strategies; physical conditioning; programs; protein expression; protein folding; public health relevance; research and development; solute; technology development; vector

DESCRIPTION (provided by applicant): A novel approach is needed to overcome the deficiencies of protein production technology and ensure the produced enzymes fold into physiologically effective three-dimensional conformations. We discovered that incorporation of novel archaeal chaperones into a host-vector system and into in vitro enzymatic reactions helps both to recover proteins from inclusion bodies and to keep enzymes operationally active in vitro. This finding opens a way for novel protein folding technology both in vivo and in vitro. We propose to develop new host-vector systems, reagents and protocols based on archaeal chaperones for protein folding in vivo and in vitro. Archaeal chaperones exhibit folding activity for a broader range of substrates because archaeal species have evolved to occupy extreme ecological niches, which pose particular challenges to the problem of protein folding. In Phase 1, we will construct several host-vector systems to demonstrate the feasibility of our approach to produce aggregation-prone proteins in their native conformations with the help of archaeal chaperones. We will also develop simple assays to accelerate technology development, streamline problem management and enable quality control for new products and protocols. A robust protein production and folding technology plays a pivotal role in biotechnology innovation. It simplifies the multiple tasks of proteomics and encourages engineering new proteins for healthcare, environmental, and industrial purposes. PUBLIC HEALTH RELEVANCE: A robust and versatile protein folding technology will impact the healthcare related R&D programs at multiple levels. Most importantly, it will facilitate high-throughput production and purification of medically important proteins. It will also bring to a new level our ability to recover and analyze effective proteins in physiological three-dimensional conformations in vitro.

描述（申请人提供）：需要一种新的方法来克服蛋白质生产技术的缺陷，并确保产生的酶折叠成生理上有效的三维构象。我们发现，将新的古细菌分子伴侣掺入宿主载体系统和体外酶促反应中有助于从包涵体中回收蛋白质并保持酶在体外的操作活性。这一发现为新的蛋白质折叠技术在体内和体外开辟了一条道路。我们建议开发新的主机载体系统，试剂和协议的基础上古细菌分子伴侣蛋白质折叠在体内和体外。古细菌分子伴侣对更广泛的底物表现出折叠活性，因为古细菌物种已经进化到占据极端的生态位，这对蛋白质折叠问题提出了特别的挑战。在第一阶段，我们将构建几个宿主载体系统，以证明我们的方法的可行性，以产生聚集倾向的蛋白质在其天然构象的帮助下的古细菌分子伴侣。我们还将开发简单的检测方法，以加速技术开发，简化问题管理，并为新产品和方案提供质量控制。强大的蛋白质生产和折叠技术在生物技术创新中发挥着关键作用。它简化了蛋白质组学的多项任务，并鼓励为医疗保健，环境和工业目的设计新蛋白质。公共卫生相关性：一种强大而通用的蛋白质折叠技术将在多个层面上影响与医疗保健相关的研发计划。最重要的是，它将促进医学重要蛋白质的高通量生产和纯化。它还将使我们在体外恢复和分析生理三维构象中有效蛋白质的能力达到一个新的水平。