Advanced bio-molecular interaction facility

1.拟支持的研究项目(目标用途)。*申请者(Boraston、Boulanger、Reynolds和Burke)在理解生物分子相互作用如何调节基本细胞过程的分子基础方面有着共同的兴趣。通过CFI和NSERC RTI之前的投资，他们已经积累了最先进的生物物理设施，用于X射线结晶学(XRC)、质谱仪(MS)、等温滴定量热法(ITC)和生化分析。这种协作的结构和生物物理设施是一个主要优势，也是加拿大生产率最高的设施之一。*拟议的研究计划是多样化的，包括微生物发病机制(Boraston、Boulanger、Reynolds和Burke)、宿主-微生物组相互作用(Reynolds)、新型生物燃料的产生(Boraston)和癌症信号(Burke、Boulanger)。几乎所有这些研究的基础是以高通量方式测量分子相互作用，特别是蛋白质-蛋白质、蛋白质-小分子和蛋白质-碳水化合物相互作用。*2.所需设备和成本(1 x Octet K2系统，138,931美元)。我们要求一种最先进的Octet生物层干涉测量系统，该系统将允许快速确定和量化生物分子相互作用。以下规格至关重要：*#1吞吐量。被提议的申请者在任何给定的时间都在研究大约50个蛋白质目标，主要目标是描述每个蛋白质可以形成的分子相互作用的特征。拥有一个能够快速表征蛋白质-蛋白质、蛋白质-碳水化合物和蛋白质-膜相互作用的平台是必不可少的，同时只需要少量的蛋白质。*#2研究复杂混合物中结合的能力。我们分析生物分子相互作用的关键是描述复杂混合物中分子相互作用的能力，包括膜混合物和生物样品。Octet K2系统能够测量极其复杂的样本中的亲和力，包括细胞裂解物。*3.与其他资金的关系。所有申请者目前都得到了NSERC探索补助金和CIHR运营补助金的支持。这笔资金不允许购买基础设施。所要求的八重奏系统将为这一研究组合提供关键支持。这些基础设施对于支持目前和未来与工业和政府合作伙伴的项目也是至关重要的，例如Zymeworks(布朗热)、阿莱克托斯(博拉斯顿、布朗热)、ARC医疗设备公司(博拉斯顿)和HHMI(伯克)。4.需要。迫切需要拟议的高度灵活的仪器来监测生物分子之间的相互作用。目前还没有能够快速表征生物分子相互作用的设备。这一要求的基础设施对于维多利亚大学的结构和生物物理研究继续取得成功至关重要。**

1. Research programs to be supported (Intended use). ***The applicants (Boraston, Boulanger, Reynolds and Burke) share a common interest in understanding the molecular basis of how bio-molecular interactions mediate fundamental cellular processes. Through previous investments from CFI and NSERC RTI they have amassed state of the art biophysical facilities for X-ray crystallography (XRC), mass spectrometry (MS), isothermal titration calorimetry (ITC), and biochemical assays. This collaborative structural and biophysical facility is a major strength, and is one of the most productive in Canada. ******The proposed research programs are diverse including microbial pathogenesis (Boraston, Boulanger, Reynolds and Burke), host-microbiome interactions (Reynolds), generation of novel biofuels (Boraston), and cancer signalling (Burke, Boulanger). Fundamental to almost all of these studies is the measurement of molecular interactions, specifically protein-protein, protein-small molecule, and protein-carbohydrate interactions in a high throughput manner.******2. Equipment requested and cost (1 x Octet K2 system at $138,931). We are requesting a state of the art Octet bio-layer interferometry system that will allow for the rapid determination and quantitation of bio-molecular interactions. The following specifications are critical:***#1 Throughput. The proposed applicants are working on ~50 protein targets at any given time, with a major goal being the characterisation of the molecular interactions each protein can form. Having a platform that allows for the rapid characterisation of protein-protein, protein-carbohydrate, and protein-membrane interactions is essential, while only needing small amounts of protein. ***#2 Capability to study binding in complex mixtures. Key to our analysis of bio-molecular interactions is the capability to characterise molecular interactions in complicated mixtures, including membrane mixtures, and biological samples. The Octet K2 system is able to measure affinities within extremely complicated samples, including cellular lysates.******3. Relationship to other funding. All of the applicants are currently supported by both NSERC Discovery grants and CIHR operating grants. This funding does not allow for infrastructure purchase. The Octet system requested will provide critical support to this research portfolio. This infrastructure will also be essential to support current and future projects with both industrial and governmental partners, for example Zymeworks (Boulanger), Alectos (Boraston, Boulanger), ARC Medical Devices (Boraston), and HHMI (Burke). ******4. Need. There is an urgent need for the proposed, highly flexible instrumentation to monitor bio-molecular interactions. There currently is no equipment that allows for the rapid characterisation of bio-molecular interactions. This requested infrastructure will be essential for the continued success of structural and biophysical research at the University of Victoria. **