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能，最终目的是在各种生物技术领域设计纳米器件。该研究计划将基础和应用科学与生物工程和合成生物学相结合。本文主要研究了海洋葡萄球菌（Staphylothermus marinus，S. marinus），一种从深海黑烟囱喷口分离出的嗜极端古细菌。S. marinus是一种严格的厌氧硫还原古菌，它利用肽作为碳源，并在高达98 ℃的温度下生长。它有一个蛋白质S层，由细长的Tetrabrachion单元组成，排列在一个3D网络中，与超热稳定蛋白酶形成蛋白质-蛋白质杂合物，称为稳定。由于其对深海火山极端条件的适应，S。marinus的特点是不寻常的稳定性。拟议的研究将侧重于调查的结构和机械特征（i）Tetrabrachion的三维组装机制，形成准晶体S层，（ii）Tetrabrachion和稳定之间的复合物形成的分子机制和（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。所有这些研究都涉及结构，生物物理和合成方法的结合，并为本科生和研究生提供各种不同的现代综合研究计划技能。自发现以来，古细菌一直受到严格的审查，因为它们对极端环境的独特适应性使它们茁壮成长。这表明生命有超嗜热起源，因此对这些生物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值，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。预期的结果是获得有关古细菌蛋白质组装体极端稳定性的机械和化学信息。了解它们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，补救和催化纳米装置等广泛领域的生物技术应用的发展提供新的见解。在未来5年内，预计3-4名研究生将在任何给定的时间从事这项研究计划。我们的研究结果将指导人工S层组件的设计和分析，对加拿大的生物技术部门产生重大影响。