Biomimetic Reagents that Promote Membrane Protein Stability (RMI)

描述（由申请人提供）：隔离在细胞膜内的蛋白质执行对健康细胞的活力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标。膜蛋白的详细结构信息是非常稀缺的，在很大程度上是由于无法产生大量的天然的，功能性的材料，是适合于结构测定。 该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。该目标将通过整合来自阿贡国家实验室（来自不同部门的多个小组）、威斯康星州大学和deCODE biostructures，Inc.的研究人员来实现。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专长的高度多学科的科学家团队。 在该方案项目中将探索的创新包括：（1）开发和实施膜蛋白结晶策略，其避免了对去污剂溶解的需要，（2）评价、设计和使用改进和替代传统去污剂的新表面活性剂，（3）选择和生产有助于稳定、纯化，和/或膜蛋白的功能形式的结晶，以及（4）用于稳定膜蛋白和产生产生结构信息的有序阵列的合成两亲性纳米材料（复合流体）的设计和应用。这些项目将得到膜蛋白生产（表达、纯化和功能表征）核心设施的支持。最初，我们将使用一组良好表征的膜蛋白作为开发和实施创新试剂，方法和方法的试验场，以解决生产稳定，功能性膜蛋白的问题，这些膜蛋白适用于纳米材料的结晶和结构测定实验。在以后的几年里，该项目中出现的技术将应用于更广泛和更具挑战性的膜蛋白靶点。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处理更苛刻，复杂和功能多样的膜蛋白和膜蛋白复合物。

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.