A protein purification platform for biochemistry and structural biology research.

本提案要求建立一个完整的系统来执行快速蛋白质液相色谱(FPLC)，以便在UQ？M建立一个完整的蛋白质表达和纯化平台。FPLC是一种实验技术，它允许在非变性和低压条件下非常有效地分离复杂的生物分子集合，以便纯化、表征、量化和鉴定初始混合物的特定化合物，通常是蛋白质、蛋白质复合体和核酸。简而言之，这种液相色谱技术是通过流动相(分析物)通过固定相(柱)洗脱，然后用检测器进行分析。这种设备将在我们的研究计划的实现中发挥核心作用，因为我们需要高纯度和特征良好的蛋白质制剂来进行生物、生化或结构研究。该部门正在进行的几个研究项目将得到该设备的支持。例如，我们将使用这种FPLC来分离可溶性蛋白质聚集体，这些蛋白质聚集体是与各种疾病相关的淀粉样纤维的前体，用于(生物)化学和生物学研究。此外，该系统将支持核磁共振对药物诱导的长QT综合征的结构研究，因为它允许纯化先前将在细菌中表达的乙醚-甲氧基相关基因(HERG)钾通道片段。将得到这一基础设施支持的其他研究项目包括了解葡萄糖-6-磷酸转移酶G6PT的结构对功能的影响，阐明铜蓝蛋白在神经系统发育中的功能，以及研究含铜的胺氧化酶的心脏保护潜力。根据这一要求，FPLC将支持的研究项目将使人们更好地了解正常和/或疾病状态下生理过程的分子基础，并最终可能导致针对不同疾病的创新治疗方法，如癌症、药物诱导的长QT综合征、蛋白质错误折叠障碍和与神经元紊乱相关的发育障碍。

The present proposal is prepared to request a complete system to perform fast protein liquid chromatography (FPLC) in oder to establish a complete platform for protein expression and purifcation at UQÀM. FPLC is an experimental technique that allows the very efficient separation, in non-denaturing and low pressure condition, of a complex ensemble of biomolecules in order to purify, characterize, quantify and identify a particular compound of the initial mixture, usually proteins, protein-complexes and nucleic acids. Briefly, this liquid chromatographic technique is supported by the elution of a mobile phase (analyte) through a stationary phase (column) following by the analysis with a detector. This equipment will play a central role for the achievement of our research programs, as we need highly-pure and well-characterized protein preparation for our biological, biochemical or structural investigations. Several research programs that are underway at the Department will be supported by this apparatus. For instance, we will use this FPLC to isolate soluble protein aggregates that are precursors of amyloid fibrils associated with various diseases for (bio)chemical and biological studies. Besides, this system will support the structural study by NMR of the drug-induced long QT syndrome by allowing the purification of fragments of the ether-à-gogo-related gene (hERG) potassium channel that will be previously expressed in bacteria. Other research projects that will be supported with this infrastructure include the understanding of the structure-to-function impact of the glucose-6-phosphate translocase G6PT, the elucidation of the function of ceruloplasmin in the nervous system development and the study of the cardioprotective potential of copper-containing amine-oxidase enzymes. The research projects that will be supported with this requested FPLC will allow a better understanding of the molecular basis of physiological processes in normal and/or disease state and could ultimately lead to innovative therapeutic approaches for different diseases, such as cancer, the drug-induced long QT syndrome, protein misfolding disorders and developmental disorders associated with neuronal disorganization.