Synthesis and Evaluation of Novel Surfactants for Membrane Protein Structural Bio

在膜蛋白的结构和功能分析过程中，蛋白质的产生一直被认为是主要的瓶颈。目前的结构基因组学研究几乎都忽略了这类重要的蛋白质，因为很难产生足够数量的功能形式的纯化样品，这是结晶实验所需要的。在生产膜蛋白样品的许多具有挑战性的步骤中，溶解的过程？将疏水膜蛋白从其天然脂质双分子层中去除，并通过使用洗涤剂胶束人工地使其与水相相容，通常是最有问题的。洗涤剂的选择被认为是一个反复试验的过程，已经被证明是非常蛋白质特异性的。目前，该领域受到表面活性剂的极大限制

Protein production has been identified as the major bottleneck in the process of analyzing structure and function of membrane proteins. Current structural genomics efforts all but ignore this important class of proteins due to the difficulty in producing sufficient quantities of purified samples in functional form, as required for crystallization experiments. Of the many challenging steps in the production of membrane protein samples, the process of solubilization ? removing the hydrophobic membrane proteins from their native lipid bilayer and artificially rendering them compatible with aqueous phases by the use of detergent micelles - is typically the most problematic. Detergent selection is known to be a trial-and-error, empirical endeavor that has proven to be very protein-specific. Currently, the field is extremely limited by surfactant sets that display only a narrow range of chemical and physical properties. Only a few classes of surfactants have been used successfully in the purification and crystallization of the approximately 60 unique membrane proteins for which structures have been deposited in the Protein Data Bank. We seek to develop efficient methods by which surfactants can be screened for their applicability to membrane protein structural biology. We will initially test these protocols using surfactants with proven utility in solubilization and maintenance of the structural and functional integrity of membrane proteins. We will then adapt these protocols for high-throughput screening in order to assess libraries of surfactants available to our team from commercial entities. These surfactants were developed for other purposes, but some of them might be applicable to this emerging field. In parallel, we will explore novel synthetic molecules that are intended to substitute for the detergents traditionally used for the solubilization of membrane proteins. Members of this latter class of molecules were specifically designed for use in membrane protein solubilization and crystallization.