Protein dynamics in iron-starved Mycobacterium tuberculosis revealed by turnover and abundance measurement using hybrid-linear ion trap-Fourier transform mass spectrometry.
To study the proteome response of Mycobacterium tuberculosis H37Rv to a change in iron level, iron-starved late-log-phase cells were diluted in fresh low- and high-iron media containing [ (15)N]-labeled asparagine as the sole nitrogen source for labeling the proteins synthesized upon dilution. We determined the relative protein abundance and protein turnover in M. tuberculosis H37Rv under these two conditions. For measurements, we used a high-resolution hybrid-linear ion trap-Fourier transform mass spectrometer coupled with nanoliquid chromatography separation. While relative protein abundance analysis shows that only 5 proteins were upregulated by high iron, 24 proteins had elevated protein turnover for the cells in the high-iron medium. This suggests that protein turnover is a sensitive parameter to assess the proteome dynamics. Cluster analysis was used to explore the interconnection of protein abundance and turnover, revealing coordination of the cellular processes of protein synthesis, degradation, and secretion that determine the abundance and allocation of a protein in the cytosol and the extracellular matrix of the cells. Further potential utility of the approach is discussed.
Rao
Prahlad K
PK
Center for Pharmaceutical Biotechnology, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60607, USA.
Rodriguez
G Marcela
GM
Smith
Issar
I
Li
Qingbo
Q
D001216:Asparagine; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D016000:Cluster Analysis; D003600:Cytosol / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005583:Fourier Analysis; D007501:Iron / Q000494:pharmacology*; D000090463:Iron Deficiencies; D013058:Mass Spectrometry / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D020543:Proteome / Q000096:biosynthesis / Q000378:metabolism; D013194:Staining and Labeling
D001426:Bacterial Proteins; D020543:Proteome; D001216:Asparagine; D007501:Iron