Characterization of bio-molecular interactions and investigation of catalytic mechanisms of bio- and chemical catalysts by micro-Isothermal Titration Calorimetry

在这项提议中，我们的目标是建立一个基础设施，用于表征生物分子相互作用、生物和化学催化剂的催化机理以及生成“智能”分子表面。我们需要一台微型等温滴定量热仪(MicroITC)。等温滴定量热法测量生物和化学反应释放的热。所要求的微型ITC的灵敏度使其能够表征具有从103到1012M-1的高到低放热和结合亲和力的反应。因此，它能够表征广泛的相互作用，例如DNA-转录因子之间的相互作用，通常具有纳摩尔范围的结合亲和力，以及蛋白质-蛋白质/肽/小分子相互作用，通常具有从低微摩尔到高微摩尔的结合亲和力。同时，这种仪器的高灵敏度将能够快速筛选适配子和小分子文库，并评估将修饰的脂质输送到细胞的效率。基于这些能力，该仪器将极大地提高这项提议申请者目前的研究能力，研究但不限于细菌对细胞壁压力的反应、抗生素耐药性、基于蛋白质的纳米管和生物细胞的形成、阿尔茨海默病的现场机制、药物的识别和开发、化学催化剂的设计和开发以及控制干细胞分化的“智能”表面的工程。这些研究领域对加拿大人的经济和健康至关重要。来自六个不同研究实验室的多达35名高素质人员(HQP)将接受这种最先进的仪器的培训。在工业环境中使用该仪器将使HQP能够很好地适应这一领域。此外，对于那些将在学术界推进其职业生涯的人来说，获得的专业知识将使他们能够推进其他研究领域，并培训更多的HQP。

In this proposal, we aim to establish an infrastructure for characterization of biomolecular interactions, catalytic mechanisms of bio and chemical catalysts and engennering of "smart" molecular surfaces. We are requesting a micro Isothermal Titration Calorimeter (microITC). Isothermal titration calorimetry measures the heat released from biological and chemical reactions. The sensitivity of the requested microITC is such that it will enable characterization of reactions that have high to low heat release and binding affinities from 103 to 1012 M-1. Hence, it enable characterization of a wide range of interactions such as those between DNA-transcription factors, which normally have binding affinities at nanomolar range, as well as protein-protein/peptide/small molecules interactions that normally have binding affinities from low micromolar to high micromolar. At the same time, the high sensitivity of such instrument would enable for quick screening of aptamers and small libraries of small molecules as well evaluation of the efficiency of delivery of modified lipids to cells . Based on these capacities this instrument will greatly enhance the current research capabilities of the applicants of this proposal to study, but not limited, to bacterial response to cell wall stress, antibiotic resistance, formation of protein-based nanotubes and biocells, mechanism of the on-set of Alzheimer's disease, identification and development of drugs and design and development of chemical catalysts and engineering of "smart" surfaces for controlling of stem cell differentiation. These research areas are essential to the economy and health of Canadians. As many as 35 high quality personnel (HQPs), coming from six different research laboratories, will be trained in this state-of-the-art instrument. Utilization of this instrument in industrial settings will enable the HQPs to be fit well in this sector. As well, for those that will advance their careers in academia, the gained expertise, will enable them to advance other research areas and train more HQPs.