Biomimetic Reagents that Promote Membrane Protein Stability (RMI)

描述(由申请人提供)：隔离在细胞膜中的蛋白质执行对健康细胞的生存至关重要的各种任务。因此，膜蛋白在疾病中起着关键作用，并代表了大多数药物靶点。膜蛋白的详细结构信息极其缺乏，这在很大程度上是由于无法生产大量适合结构确定的天然功能材料。 该计划项目的目标是开发和实施促进膜蛋白稳定性和结晶的仿生试剂。这一目标将通过将来自阿贡国家实验室(来自不同部门的多个小组)、威斯康星大学和DeCODE BioStructures，Inc.的研究人员整合到一个高度多学科的科学家团队中来实现，这些科学家在生物化学、合成化学、生物物理学、免疫学和材料科学方面具有互补的专业知识。 该计划项目将探索的创新包括：(1)开发和实施避免洗涤剂增溶需要的膜蛋白结晶策略，(2)评估、设计和使用改进和取代传统洗涤剂的新型表面活性剂，(3)选择和生产有助于稳定、纯化和/或结晶功能形式的膜蛋白的亲和剂，以及(4)用于稳定膜蛋白的合成两亲性纳米材料(复合液)的设计和应用，以及产生结构信息的有序阵列的生产。这些项目将得到膜蛋白生产(表达、纯化和功能鉴定)核心设施的支持。首先，我们将使用一组特性良好的膜蛋白作为试验场，开发和实施创新的试剂、方法和途径，以解决生产稳定、功能良好的膜蛋白的问题，这些膜蛋白适合于纳米材料的结晶和结构确定实验。在接下来的几年里，该项目产生的技术将应用于更广泛、更具挑战性的膜蛋白靶标，并进行调整。从早期的成功和失败中获得的见解应该允许一种智能和有效的方法来开发一套要求更高、更复杂、功能更多样化的膜蛋白和膜蛋白复合体。

DESCRIPTION (provided by applicant): Proteins sequestered within cellular membranes perform a variety of tasks that are critical for the viability of healthy cells. As such, membrane proteins play pivotal roles in disease and represent the majority of drug targets. Detailed structural information for membrane proteins is exceedingly scarce, due in large part to the inability to produce quantities of native, functional material that is suitable for structure determination. The objective of this program project is to develop and implement biomimetic reagents that promote stability and crystallization of membrane proteins. This objective will be met by integrating investigators from Argonne National Laboratory (multiple groups from different divisions), the University of Wisconsin, and deCODE biostructures, Inc. into a highly multidisciplinary team of scientists with complementary expertise in biochemistry, synthetic chemistry, biophysics, immunology, and materials science. The innovations to be explored within this program project include: (1) development and implementation of a membrane protein crystallization strategy that circumvents the need for detergent solubilization, (2) evaluation, design, and use of new surfactants that improve upon and replace traditional detergents, (3) selection and production of affinity reagents that assist in the stabilization, purification, and/or crystallization of functional forms of membrane proteins, and (4) the design and application of synthetic amphiphilic nanomaterials (complex fluids) for the stabilization of membrane proteins and the production of ordered arrays that yield structural information. These projects will be supported by a core facility for membrane protein production (expression, purification, and functional characterization). Initially, we will use a set of well-characterized membrane proteins as a testing ground for the development and implementation of innovative reagents, methods, and approaches to the problem of producing stable, functional membrane proteins that are suitable for crystallization and structure determination experiments in nanomaterials. In later years, techniques emerging from the project will be applied to and adjusted for a broader and more challenging range of membrane protein targets. The insights gained from early successes and failures should allow an intelligent and effective approach to a more demanding, intricate, and functionally diverse set of membrane proteins and membrane protein complexes.