Biophysical Society Thematic Meeting - November 16-20, 2015

Biophysics in the Understanding, Diagnosis, and Treatment of Infectious Diseases Speaker Abstracts

Targeting the Membrane Proteome of mTB for Structure based Approaches to Function Robert M. Stroud, Oren Rosenberg, Jonny Leano, Hemant Kumar, Karolina Kaminska, Yaneth Robles. Department of Biochemistry, University of California in San Francisco, USA. There are currently 803 transmembrane proteins in the mTB genome. There is no atomic structure for even one of these proteins, yet they govern the signaling, entry and exit from the cell. These proteins govern functions that are variously important, sometimes essential for the viability and virulence of mTB. They govern the import of nutrients and secretion systems for the export of virulence factors, and proteins that are adapted to export drugs used to treat tuberculosis. We describe a system to select among the integral membrane proteome of mTB with the end goal of determining their structure at atomic level, sufficient to determine interaction with fragments of molecules and compounds from libraries intended to block critical and essential functions in mTB alone, avoiding interference with the human host by developing selectivity. A high throughput cloning and expression for a selected and focused set of mTB membrane proteins are presented. Examples illustrate how atomic structures of integral membrane protein can be pursued and determined by X-ray crystallography, and the prospects of understanding mechanisms of them by other means. These include the generation of antibody Fab fragments from phage displayed libraries both as tools to modulate activity, and as structural aids to crystallization of electron cryo-microscopy. Illustration of recent applications of these methods and what can be learned from these approaches are presented. http://www.msg.ucsf.edu/stroud/index.htm

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