Automated Protein Crystal Growth in Lipidic Cubic Phase

Nollert-Von Specht, Peter

Abstract

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 Applications

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.