pr400334k_si_004.xlsx (107.8 kB)
Comparison of the Membrane Proteome of Virulent Mycobacterium tuberculosis and the Attenuated Mycobacterium bovis BCG Vaccine Strain by Label-Free Quantitative Proteomics
dataset
posted on 2013-12-06, 00:00 authored by Harsha
P. Gunawardena, Meghan E. Feltcher, John A. Wrobel, Sheng Gu, Miriam Braunstein, Xian ChenThe Mycobacterium tuberculosis membrane is rich
in antigens that are potential targets for diagnostics and the development
of new vaccines. To better understand the mechanisms underlying MTB
virulence and identify new targets for therapeutic intervention, we
investigated the differential composition of membrane proteomes between
virulent M. tuberculosis H37Rv (MTB) and the Mycobacterium bovis BCG vaccine strain. To compare the membrane
proteomes, we used LC–MS/MS analysis in combination with label-free
quantitative proteomics, utilizing the area under the curve of the
extracted ion chromatograms of peptides obtained from m/z and retention time alignment of MS1 features.
With this approach, we obtained relative abundance ratios for 2203
identified membrane-associated proteins in high confidence. Of these
proteins, 294 showed statistically significant differences of at least
two fold in relative abundance between MTB and BCG membrane fractions.
Our comparative analysis detected several proteins associated with
known genomic regions of difference between MTB and BCG as being absent,
which validated the accuracy of our approach. In further support of
our label-free quantitative data, we verified select protein differences
by immunoblotting. To our knowledge, we have generated the first comprehensive
and high-coverage profile of comparative membrane proteome changes
between virulent MTB and its attenuated relative BCG, which helps
elucidate the proteomic basis of the intrinsic virulence of the MTB
pathogen.