Methodology

The structure determination of membrane proteins is significantly lagging behind water-soluble proteins. One major impediment is the lack of strategies for obtaining well-diffracting membrane protein crystals, although difficulties in expression and functional isolation of membrane proteins also hamper the process. Faced with these challenges, the MPID Center combines the use of various technologies that has been developed by current PSI centers and applies this experience to the development of new strategies that may allow higher throughput of membrane protein structure determination.

The new strategies will minimize the bottlenecks for membrane protein structure determination at all levels of the structure determination pipeline. Specifically, we will implement and optimize systems for high throughput expression of functional membrane proteins, establish efficient semi-automated protocols for rapid and mild solubilization and purification of membrane proteins, construct biochemical and biophysical characterization related to the functional/structural integrity of the membrane proteins, develop a rationale design of membrane protein crystallization, and a suite of bioinformatics tools to computationally predict membrane proteins’ structures.