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: 7940175
• 负责人: FRANK Thomson ROBB
• 金额: $ 9.38万
• 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/7940175
• 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（由申请人提供）：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有效的三维构象。我们发现，将新颖的古细菌伴侣掺入宿主矢量系统中，并将其掺入体外酶促反应中，有助于从包含体中恢复蛋白质，并使酶在体外保持活性。这一发现为体内和体外的新型蛋白质折叠技术打开了一种方法。我们建议开发新的宿主矢量系统，试剂和方案，基于古细菌伴侣，用于体内和体外蛋白质折叠。古细菌伴侣在更广泛的底物上表现出折叠活性，因为古细菌已经进化为占据极端的生态壁ni，这对蛋白质折叠的问题构成了特别的挑战。在第1阶段，我们将构建几个宿主矢量系统，以证明我们在借助古细菌伴侣的帮助，在其天然构型中产生聚集的蛋白质的可行性。我们还将开发简单的测定，以加速技术开发，简化问题管理并实现新产品和协议的质量控制。强大的蛋白质生产和折叠技术在生物技术创新中起关键作用。它简化了蛋白质组学的多种任务，并鼓励工程新蛋白质用于医疗保健，环境和工业目的。公共卫生相关性：强大而多功能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，它将促进医学上重要的蛋白质的高通量产生和纯化。这也将使我们在体外生理三维构象中恢复和分析有效蛋白质的有效蛋白质的新水平。