Biophysical Society Thematic Meeting

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.