Producing medically important human membrane proteins for structural studies.

Abstract

Integral membrane proteins are essential for many activities within a cell but the structure of only few of these proteins have been solved. Highly purified amounts of proteins are required for crystallization and structural studies thus signifying a need to improve techniques to achieve this purpose. A total of five different membrane proteins (Ergic1, Ergic2, Ergic3, Mrs2 and Stim1) were studied for the testing of new purification strategies. This project aims to produce high quantities of medically relevant human membrane proteins suitable for crystallization using strategies like screening different vectors, cell strains and induction systems. The use of a double affinity tag purification system was used to increase the purity and expression of proteins in different Escherichia coli host cells. The membrane protein, Ergic2, was successfully expressed and purified. The success of this purification strategy will pave the way for large scale production of highly purified proteins that can be used for crystallization studies.