SIG Program: AKTAxpress FPLC System for Protein Purification

描述(由申请人提供)：AKT Axpress FPLC蛋白质纯化系统1)项目摘要/摘要蛋白质纯化能力是结构生物学、蛋白质设计和蛋白质科学各个方面的基础。当必须处理大量样品和优化多步骤纯化方案时，高通量自动蛋白质纯化可以极大地提高研究生产率。加州理工大学的三个领先的蛋白质研究实验室(Arnold，Rees，Tirrell)正在请求支持，以购买AKTAxpress FPLC(快速蛋白质液相色谱)系统。该仪器能够在无人值守的情况下执行全自动、多步骤纯化，从而为结晶学和生化蛋白质表征等应用提供尽可能高的纯度和高产率。模块化的设置允许不同的纯化方案在不同的模块上同时运行，使三个研究小组能够在不相互干扰的情况下纯化他们的蛋白质。这台仪器将消除我们研究工作的一个非常重要的瓶颈：蛋白质纯化。目前的净化系统上的时间已经排满，需要大量的研究人员投入，而AKTAxpress将把这一投入降至最低。模块化设置还允许在需要时扩展功能(增加了新的净化‘模块’，总共多达12个)。该仪器将支持对细胞色素P450功能的结构基础的研究，并将为合成人类代谢物和药物铅多样化(Arnold)、膜蛋白质晶体结构分析(REES)和通过掺入非天然氨基酸(Tirrell)来工程蛋白质设计新的P450。细胞色素P450酶是人类药物代谢的关键因素，了解它们的功能对于开发安全、有效的治疗药物至关重要。这些酶可用于合成真正的代谢物，用于临床测试(例如，毒性或增强治疗功能)。整体膜蛋白是大多数常用处方药的靶标，但其结构分析仍处于初级阶段。通过结合非天然氨基酸进行蛋白质工程为基于蛋白质的治疗和诊断以及新的研究工具提供了令人兴奋的新机会。 公共卫生相关性：由这项仪器拨款支持的研究将有助于更好地了解药物代谢、药物-药物相互作用和药物作用机制。蛋白质纯化是结构生物学、蛋白质设计和蛋白质科学方方面面的基本工具。蛋白质纯化目前是所有三个需要这种仪器的小组的瓶颈，他们的研究生产率将通过所要求的蛋白质纯化系统得到显著提高。

DESCRIPTION (provided by applicant): AKTAxpress FPLC System for Protein Purification 1) Project Summary/Abstract Protein purification capabilities underlie structural biology, protein design, and all aspects of protein science. High-throughput automated protein purification can greatly enhance research productivity when many samples must be handled and multi-step purification protocols optimized. Three leading protein research laboratories at Caltech (Arnold, Rees, Tirrell) are requesting support to purchase an AKTAxpress FPLC (Fast Protein Liquid Chromatography) system. This instrument is capable of performing fully- automated, multi-step purifications in an unattended fashion, resulting in highest possible purity and high yields for applications such as crystallography and biochemical protein characterization. The modular setup allows different purification protocols to be run simultaneously on different modules, enabling the three research groups to purify their proteins without interfering with each other. This instrument will remove a very significant bottleneck to our research efforts: protein purification. Time on current purification systems is fully booked, and requires significant researcher input that will be minimized with the AKTAxpress. The modular setup also allows expansion of capabilities when needed (with addition of new purification 'modules', for a total of up to 12). This instrument will support research into the structural basis of cytochrome P450 function and engineering new P450s for synthesis of human metabolites and drug lead diversification (Arnold), membrane protein crystallographic structure analysis (Rees), and engineering proteins through incorporation of nonnatural amino acids (Tirrell). Cytochrome P450 enzymes are key players in human drug metabolism, and understanding their function is critical to development of safe, effective therapeutics. These enzymes can be used to synthesize authentic metabolites for clinical testing (e.g. for toxicity or enhanced therapeutic function). Integral membrane proteins are targets of most popularly prescribed drugs, yet their structural analysis is still in its infancy. Protein engineering via incorporation of nonnatural amino acids offers exciting new opportunities for protein-based therapeutics and diagnostics as well as new research tools. PUBLIC HEALTH RELEVANCE: The research supported by this instrumentation grant will lead to a better understanding of drug metabolism, drug-drug interactions, and mechanisms of drug action. Protein purification is a fundamental tool in structural biology, protein design, and all aspects of protein science. Protein purification is currently a bottleneck for all three groups requesting this instrumentation, and their research productivity will be significantly enhanced by the requested protein purification system.