TY - JOUR

T1 - A chemical proteomics approach to profiling the ATP-binding proteome of Mycobacterium tuberculosis

AU - Wolfe, Lisa M

AU - Veeraraghavan, Usha

AU - Idicula-Thomas, Susan

AU - Schürer, Stephan

AU - Wennerberg, Krister

AU - Reynolds, Robert

AU - Besra, Gurdyal S

AU - Dobos, Karen M

PY - 2013/6

Y1 - 2013/6

N2 - Tuberculosis, caused by Mycobacterium tuberculosis, remains one of the leading causes of death worldwide despite extensive research, directly observed therapy using multidrug regimens, and the widespread use of a vaccine. The majority of patients harbor the bacterium in a state of metabolic dormancy. New drugs with novel modes of action are needed to target essential metabolic pathways in M. tuberculosis; ATP-competitive enzyme inhibitors are one such class. Previous screening efforts for ATP-competitive enzyme inhibitors identified several classes of lead compounds that demonstrated potent anti-mycobacterial efficacy as well as tolerable levels of toxicity in cell culture. In this report, a probe-based chemoproteomic approach was used to selectively profile the M. tuberculosis ATP-binding proteome in normally growing and hypoxic M. tuberculosis. From these studies, 122 ATP-binding proteins were identified in either metabolic state, and roughly 60% of these are reported to be essential for survival in vitro. These data are available through ProteomeXchange with identifier PXD000141. Protein families vital to the survival of the tubercle bacillus during hypoxia emerged from our studies. Specifically, along with members of the DosR regulon, several proteins involved in energy metabolism (Icl/Rv0468 and Mdh/Rv1240) and lipid biosynthesis (UmaA/Rv0469, DesA1/Rv0824c, and DesA2/Rv1094) were found to be differentially abundant in hypoxic versus normal growing cultures. These pathways represent a subset of proteins that may be relevant therapeutic targets for development of novel ATP-competitive antibiotics.

AB - Tuberculosis, caused by Mycobacterium tuberculosis, remains one of the leading causes of death worldwide despite extensive research, directly observed therapy using multidrug regimens, and the widespread use of a vaccine. The majority of patients harbor the bacterium in a state of metabolic dormancy. New drugs with novel modes of action are needed to target essential metabolic pathways in M. tuberculosis; ATP-competitive enzyme inhibitors are one such class. Previous screening efforts for ATP-competitive enzyme inhibitors identified several classes of lead compounds that demonstrated potent anti-mycobacterial efficacy as well as tolerable levels of toxicity in cell culture. In this report, a probe-based chemoproteomic approach was used to selectively profile the M. tuberculosis ATP-binding proteome in normally growing and hypoxic M. tuberculosis. From these studies, 122 ATP-binding proteins were identified in either metabolic state, and roughly 60% of these are reported to be essential for survival in vitro. These data are available through ProteomeXchange with identifier PXD000141. Protein families vital to the survival of the tubercle bacillus during hypoxia emerged from our studies. Specifically, along with members of the DosR regulon, several proteins involved in energy metabolism (Icl/Rv0468 and Mdh/Rv1240) and lipid biosynthesis (UmaA/Rv0469, DesA1/Rv0824c, and DesA2/Rv1094) were found to be differentially abundant in hypoxic versus normal growing cultures. These pathways represent a subset of proteins that may be relevant therapeutic targets for development of novel ATP-competitive antibiotics.

KW - Adenosine Triphosphate/antagonists & inhibitors

KW - Antitubercular Agents/chemistry

KW - Bacterial Proteins/antagonists & inhibitors

KW - Binding, Competitive

KW - Carrier Proteins/antagonists & inhibitors

KW - Culture Media

KW - Gene Expression Regulation, Bacterial/drug effects

KW - Isocitrate Lyase/genetics

KW - Mycobacterium tuberculosis/drug effects

KW - Oxygen/metabolism

KW - Peptides/chemistry

KW - Protein Binding

KW - Protein Interaction Mapping

KW - Protein Isoforms/antagonists & inhibitors

KW - Protein Kinases/genetics

KW - Proteome/antagonists & inhibitors

KW - Proteomics/methods

KW - Signal Transduction

U2 - 10.1074/mcp.M112.025635

DO - 10.1074/mcp.M112.025635

M3 - Journal article

C2 - 23462205

VL - 12

SP - 1644

EP - 1660

JO - Molecular and Cellular Proteomics

JF - Molecular and Cellular Proteomics

SN - 1535-9476

IS - 6

ER -