New chaperones for folding and assembly of proteins

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，并确保生产的酶折叠成生理上有效的三维构象。我们发现，将新的古细菌伴侣结合到宿主-载体系统和体外酶促反应中，有助于从包涵体中恢复蛋白质，并保持酶在体外的操作活性。这一发现为体内和体外的新型蛋白质折叠技术开辟了一条道路。我们建议在古细菌伴侣蛋白的基础上开发新的宿主载体系统、试剂和方案，用于体内和体外蛋白质折叠。古细菌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来证明我们的方法在古细菌伴侣的帮助下以其天然构象生产易于聚集的蛋白质的可行性。我们还将开发简单的分析方法，以加速技术开发，简化问题管理，并使新产品和新协议的质量控制成为可能。强大的蛋白质生产和折叠技术在生物技术创新中起着关键作用。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的蛋白质。公共卫生相关性：一项强大而通用的蛋白质折叠技术将在多个层面上影响医疗保健相关的研发项目。最重要的是，它将促进高通量生产和纯化医学上重要的蛋白质。它还将把我们在体外生理三维构象中恢复和分析有效蛋白质的能力提升到一个新的水平。

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.