Critical Replacement of a Differential Scanning Calorimeter for Protein Studies

蛋白质研究中差示扫描量热计的关键替代品

• 批准号: RTI-2023-00360
• 负责人: Bearne, Stephen
• 金额: $ 10.93万
• 依托单位: Dalhousie University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 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=744249
• 关键词: Critical; Replacement; Differential; Scanning; Calorimeter

Differential scanning calorimetry (DSC) is a powerful experimental technique for determining the thermodynamic properties accompanying protein-protein binding, protein unfolding, ligand binding, and the stability of biomolecular assemblies. This grant will replace a malfunctioning, unrepairable differential scanning calorimeter, to permit completion of on-going experiments and publication of the results, and enable new experiments for the characterization of biomacromolecules to be conducted at Dalhousie University. The fundamental information gained using DSC is essential for understanding enthalpic and entropic contributions to protein stability and intermolecular interactions, and for re-engineering and producing new proteins with desirable properties. The ability of DSC to monitor the unfolding, phase transitions, and degree of oligomerization of proteins, polynucleotides, lipid assemblies, and other biomolecular structures makes it extremely useful for the study of biomolecules. All 5 NSERC-funded applicants and their 28 trainees, ranging from undergraduates to postdoctoral fellows and professional research associates, with interests spanning the life and biomedical sciences to fundamental and applied materials chemistry, urgently need to regain access to this valuable biophysical technique. The instrument will permit optimum calorimetric characterization of proteins such as spider silks, antifreeze proteins, hydrophobins, hemolymph proteins, and enzymes, as well as maltodextrin-based particles, with the potential for new biomedical, biotechnological, and ecological applications. Specifically, applicants and trainees will be able to (1) evaluate the role if active site residues to enzyme stability, (2) assess the effect of interdigitating loops on the stability of enzyme oligomers, (3) determine the effects of protein mixing or mutation on the thermal stability of silk fusion proteins and their oligomers, (4) characterize the interactions of antifreeze proteins with cell membranes, (5) develop a novel approach using the hemolymph of blue mussels for environmental monitoring, (6) evaluate the thermal properties of semicrystalline particles used in drug delivery, and (7) assess the stability of assemblies of hydrophobins. No instrument with the sensitivity and appropriate configuration for analyzing dilute protein solutions is available to the applicants either at Dalhousie or nearby institutions, making acquisition essential to support and enhance their research and training programs. The instrument will be installed in Dalhousie's Biophysical Characterization Facility, providing ready access to the co-applicants and to many other researchers at Dalhousie and surrounding institutions. The applicants support a diverse and inclusive training environment where trainees will gain both hands-on experience and valuable expertise with this important biophysical technique that is often employed in pharma and biotechnology industries.

差示扫描量热法（DSC）是一种强大的实验技术，用于确定伴随蛋白质-蛋白质结合，蛋白质解折叠，配体结合和生物分子组装体稳定性的热力学性质。这笔赠款将取代一个故障，不可修复的差示扫描量热仪，以允许正在进行的实验和结果的出版完成，并使生物大分子的表征新的实验将在达尔豪西大学进行。使用DSC获得的基本信息对于理解蛋白质稳定性和分子间相互作用的热力学和熵贡献以及重新设计和生产具有所需特性的新蛋白质是必不可少的。DSC监测蛋白质、多核苷酸、脂质组装体和其他生物分子结构的解折叠、相变和寡聚化程度的能力使其对于生物分子的研究非常有用。所有5名NSERC资助的申请人及其28名学员，从本科生到博士后研究员和专业研究人员，其兴趣涵盖生命和生物医学科学到基础和应用材料化学，迫切需要重新获得这种宝贵的生物物理技术。该仪器将允许蛋白质，如蜘蛛丝，抗冻蛋白，疏水蛋白，血淋巴蛋白和酶，以及麦芽糖糊精为基础的颗粒的最佳量热表征，具有新的生物医学，生物技术和生态应用的潜力。具体而言，申请人和受训者将能够（1）评估活性位点残基对酶稳定性的作用，（2）评估交叉指状环对酶寡聚体稳定性的影响，（3）确定蛋白质混合或突变对丝融合蛋白及其寡聚体热稳定性的影响，（4）表征抗冻蛋白与细胞膜的相互作用，（5）开发一种使用蓝贻贝血淋巴进行环境监测的新方法，（6）评估用于药物递送的半结晶颗粒的热性质，以及（7）评估疏水蛋白组装体的稳定性。申请人在达尔豪西或附近的机构都没有具有分析稀释蛋白质溶液的灵敏度和适当配置的仪器，这使得收购对于支持和加强他们的研究和培训计划至关重要。该仪器将安装在达尔豪西的生物物理表征设施，为共同申请人和达尔豪西及周边机构的许多其他研究人员提供方便。申请人支持多元化和包容性的培训环境，学员将获得实践经验和宝贵的专业知识，这一重要的生物物理技术通常用于制药和生物技术行业。