Automated Protein Crystal Growth in Lipidic Cubic Phase

从蛋白质的三维结构中获得的信息在药物开发中越来越重要。使用三维x射线晶体学确定的结构在抗艾滋病和抗癌药物的开发中至关重要。膜蛋白构成了基因的很大一部分，并且是市场上大约60%的药物的靶标。膜蛋白三维结构的测定远远落后于可溶性蛋白结构的测定。最近开发的一种膜蛋白晶体生长技术是使用脂质立方相（LCP）材料，其中膜蛋白在类似天然脂质双分子层的微环境中结晶。处理这些材料的一个困难是lcp -蛋白质混合物的极高粘度。使用注射器分配LCP材料的传统方法已经开发出来，但是从注射器中自动填充和分配这种高粘度材料一直具有挑战性，并且基于LCP的晶体生长的新方法正在开发中，需要将LCP无接触地分配到基板上。该项目的目标是开发一种创新的方法，将LCP无接触地分配到底物上，使膜蛋白晶体生长以高通量的方式自动化。建议的商业应用：我们建议的项目将有助于膜蛋白结构的测定。任何能够加速确定这些结构的技术在开发新药方面都具有重要价值。Emerald是UCSF开发的LCP技术的独家授权方。Emerald BioStructures聘请了三位具有膜蛋白经验的晶体学家Craig Behnke、Thomas Richter和Peter nolert，后者是加州大学旧金山分校LCP技术的开发者之一。

Information derived from the three-dimensional structure of proteins has been of increasing importance in the development of pharmaceuticals. Structures determined using 3-D X-ray crystallography have been vital in the development of anti-AIDS and anti-cancer agents. Membrane proteins constitute a large fraction of genes, and are the target of approximately 60% of the drugs on the market. Determinations of the 3-D structures of membrane proteins have lagged far behind the determination of structures of soluble proteins. One technique recently developed for membrane protein crystal growth has been the use of lipidic cubic phase (LCP) materials in which the membrane protein crystallizes in a microenvironment resembling native lipid bilayer. One difficulty in working with these materials is the extreme viscosity of the LCP-protein mixture. Conventional methods of dispensing LCP materials using a syringe have been developed, but automating filling and dispensing of this high-viscosity material from a syringe has been challenging, and new methods for LCP-based crystal growth under development require contact-free dispensation of LCP onto substrate. The goal of this project is to develop an innovative method for contact-free dispensation of LCP onto a substrate, allowing membrane protein crystal growth to be automated in a high-throughput manner. PROPOSED COMMERCIAL APPLICATION: Our proposed project will facilitate the determination of membrane protein structures. Any technology developed to speed the determination of these structures would be of great value in developing novel pharmacuticafs. Emerald is the exclusive licensee of the LCP technology developed at UCSF. Emerald BioStructures has employed three crystallographers with membrane protein experience, Craig Behnke, Thomas Richter, and Peter Nollert, one of the developers of LCP technology at UCSF.