Structure-Property relationship of S-layer protein assemblies at extremophilic archea bacteria

极端古细菌S层蛋白组装体的结构-性质关系

• 批准号: RGPIN-2018-04970
• 负责人: Stetefeld, Jörg
• 金额: $ 6.12万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748106
• 关键词: Structure; Property; relationship; layer; protein

Nature of the WorkThe long term objective of my research program is to investigate the extreme stability and unique properties of archaea Surface layer (S-layer) components for the ultimate purpose to engineer nano-devices in a variety of biotechnological fields. The research program combines Basic -and Applied Science with Bio-engineering and Synthetic Biology. We focus our attention on the structure-property relationship of the unique S-layer protein assembly from Staphylothermus marinus (S. marinus), an extremophillic archaea bacterium isolated from deep sea black smoker vents. S. marinus is a strict anaerobic sulfur reducing archaeon that utilizes peptides as a carbon source and grows at temperatures up to 98C. It has a proteinaceous S-layer consisting of elongated Tetrabrachion units arranged in a 3D-network that forms a protein-protein heterocomplex with the hyperthermostable protease, called STABLE. As a consequence of its adaptation to the extreme conditions at the deep sea volcano, the S-layer proteins of S. marinus are characterized by an unusual stability. The proposed research will focus on investigating structural and mechanistic features of (i) the 3D-assembly mechanism of Tetrabrachion to form a quasi-crystalline S-layer, (ii) the molecular mechanisms of complex formation between Tetrabrachion and STABLE and (iii) the extreme stability of S-layer components as a foundation for bio-engineering and synthetic biology. All these studies involve a combination of structural, biophysical and synthetic approaches and offer undergraduate and graduate students a great variety of different skills in modern integrated research programs. ImportanceSince their discovery, archaea have been under intense scrutiny due to their unique adaptations to the extreme environments in which they thrive. It is suggested that life has hyperthermophilic origins, and thus the study of these organisms provides insight into the biology of early life forms. In addition, understanding the molecular mechanisms of their adaptation to the extreme conditions (pH, Temperature, pressure, salinity, ionic strength) makes these microorganisms a perfect research tool for biotechnological developments. Outcome and BenefitThe expected outcome is to obtain mechanistic and chemical information about the extreme stability of archaeal protein assemblies. Understanding the molecular basis of their unique adaptation, will open new perspectives on our understanding of the origin of life and will provide new insights into development of biotechnological applications in the broad fields of monitoring, remediation and catalytic nano-devices. Over the next 5 years, it is expected that 3-4 graduate students be engaged in this research program at any given time. Our findings will guide the design and analysis of artificial S-layer assemblies with great impact on Canada's biotech sector.

工作性质我的研究计划的长期目标是研究古细菌表面层（S 层）组件的极端稳定性和独特性质，最终目的是在各种生物技术领域设计纳米器件。该研究计划将基础科学和应用科学与生物工程和合成生物学相结合。我们将注意力集中在来自海洋葡萄球菌 (S. marinus) 的独特 S 层蛋白质组装的结构-性质关系上，海洋葡萄球菌是一种从深海黑烟囱中分离出来的极端嗜热古细菌。 S. marinus 是一种严格厌氧的硫还原古菌，利用肽作为碳源，并在高达 98°C 的温度下生长。它有一个蛋白质 S 层，由排列在 3D 网络中的细长 Tetrabrachion 单元组成，与超热稳定蛋白酶（称为 STABLE）形成蛋白质-蛋白质杂复合物。由于适应了深海火山的极端条件，S. marinus 的 S 层蛋白具有不同寻常的稳定性。拟议的研究将重点研究以下结构和机械特征：(i) Tetrabrachion 形成准晶体 S 层的 3D 组装机制，(ii) Tetrabrachion 和 STABLE 之间复合物形成的分子机制，以及 (iii) S 层组件的极端稳定性，作为生物工程和合成生物学的基础。所有这些研究都结合了结构、生物物理和合成方法，为本科生和研究生提供了现代综合研究项目中各种不同的技能。重要性自从被发现以来，古细菌因其对生长的极端环境的独特适应而受到密切关注。有人认为生命具有超嗜热起源，因此对这些生物体的研究提供了对早期生命形式生物学的深入了解。此外，了解它们适应极端条件（pH、温度、压力、盐度、离子强度）的分子机制使这些微生物成为生物技术发展的完美研究工具。结果和效益预期的结果是获得有关古菌蛋白质组装体的极端稳定性的机械和化学信息。了解其独特适应的分子基础，将为我们理解生命起源开辟新的视角，并将为监测、修复和催化纳米设备等广泛领域的生物技术应用的发展提供新的见解。未来 5 年内，预计随时有 3-4 名研究生参与该研究项目。我们的研究结果将指导人工 S 层组件的设计和分析，对加拿大生物技术领域产生重大影响。