The Transmembrane Protein Center (TMPC) was funded from July 2010 - June 2015 through the Protein Structure Initiative (PSI) with the funding from the National Institute of General Medical Sciences (NIGMS), one of the National Institutes for Health (NIH). The goal of the TMPC was to apply technologies developed under the aegis of the PSI:Biology Network to problems of interest to the community of biologists and biochemists who investigate the role of membrane proteins in human health and disease.
The collective experience of our research team suggests that application of many techniques and efficient screening through many types of expression, stabilization and purification protocols will be needed to expand the chances for success with the broad range of membrane proteins expected from participation in the PSI:Biology Network.
Our second goal was to establish an efficient membrane protein production pipeline to advance understanding of the structure and function of membrane proteins. An overview of our approach is provided at the right. We included capability to create protein variants, to refine screening based on functional and biophysical characterizations, and to sort between multiple expression pathways to find the best way produce a membrane protein of interest. Emphasis on small-scale purification screening and integration of automation into our efforts contributes to our efforts. Options for crystallization screening, access to synchrotrons, and improvements in software used to solve structures also provides resources and skills necessary to achieve PSI:Biology goals.
TMPC evaluated new target nominations and actively sought collaborators who could contribute biological assays as an integral part of screening process. Toward this end, TMPC provided cell-free translation plasmids and cloning protocols to research groups currently outside of the PSI:Biology Network. Our initial stages of collaboration involved production of sequence-verified expression plasmids, small-scale expression testing in wheat germ cell-free translation, and return of samples to the collaborator for functional studies. Best performing candidates from these initial screening studies were be adopted for additional scale-up studies depending on resources available.