A microscale thermophoresis instrument for characterizing molecular interactions

最近，微尺度热电泳法(MST)已经成为一种强大的技术，用于表征蛋白质、DNA和脂质等各种分子之间的相互作用。MST依赖于热渗透现象，这是当置于温度梯度中时分子的运动。与其他生物物理技术相比，MST有几个优势：i)它非常敏感，需要的样本很少；ii)分析可以在复杂的缓冲液或样本中进行，如血清或细胞裂解物；iii)数据收集简单、快速，在一次性毛细管中进行，消除了污染，并允许进行有效的用户培训。这笔赠款将为NSERC资助的7名共同申请者和他们研究小组中的约43名高素质人员提供微型温热导入(MST)仪器。共同申请人研究范围广泛的生物技术和生物医学问题，并将能够完成特定的目标，如1)表征疏水蛋白与蛋白质和生物材料的相互作用，2)确定黑素细胞发育的调节机制，3)分子拥挤对苦杏仁酸消旋酶抑制剂的影响，4)调查KapB在调节细胞质加工机构中的作用，5)深入了解蛋白质与细胞囊泡相互作用诱导膜融合的速度，6)开发苦味受体的抑制剂，7)研究修饰的p53蛋白与细胞结合伙伴的相互作用，以及8)小分子如何影响代谢途径。 申请者没有类似的设备，因此必须购买一台MST仪器来支持申请者的研究计划。该仪器将设在达尔豪西大学蛋白质组学核心设施内，便于海洋区域的其他研究人员使用该仪器。一名全职工作人员将负责该仪器，并为有兴趣使用该仪器的HQP提供培训。这将使学员学会进行MST实验并解释结果，这将是他们未来职业生涯中的一项宝贵技能。

Recently, microscale thermophoresis (MST) has emerged as a powerful technique that is used to characterize interactions between a variety of molecules such as proteins, DNA and lipids. MST relies on the phenomenon of thermophoresis, which is the movement of molecules when placed in a temperature gradient. There are several advantages that MST has over other biophysical techniques: i) It is extremely sensitive and requires very little sample; ii) Analysis can take place in complex buffers or samples such blood serum or cell lysate; iii) Data collection is simple, fast and takes place in disposable capillaries, eliminating contamination and allowing for efficient user training. This grant will provide ready access of a microscale thermophoresis (MST) instrument to the 7 NSERC-funded co-applicants and the ~43 highly qualified personnel in their research groups. The co-applicants study a wide range of biotechnical and biomedical problems and will be able to complete specific goals such as 1) characterize hydrophobin interactions with proteins and biomaterials, 2) determine the regulatory mechanisms of melanocyte development, 3) characterize the effect of molecular crowding on mandalate racemease inhibitors 4) investigate the involvement of KapB in the regulation of cytoplasmic processing bodies, 5) gain insight to how FAST proteins interact with cellular vesicles to induce membrane fusion, 6) develop inhibitors to bitter taste receptors, 7) study the interactions of modified p53 proteins with cellular binding partners, and 8) investigate how small molecules can influence metabolic pathways. No similar equipment is available to the applicants, making it essential to acquire a MST instrument to support the applicants’ research programs. The instrument will be located in the Dalhousie University Proteomics Core Facility, allowing for easy access of the instrument to other researchers in the Maritime region. A full time staff member will be responsible for the instrument and providing training to HQP interested in using it. This will allow trainees to learn to conduct MST experiments and interpret results, which will be a valuable skillset in their future careers.