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Identification of acidic, low molecular mass proteins of Mycobacterium tuberculosis strain H37Rv by matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry

Item Type:Article
Title:Identification of acidic, low molecular mass proteins of Mycobacterium tuberculosis strain H37Rv by matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry
Creators Name:Mattow, J., Jungblut, P.R., Mueller, E.C. and Kaufmann, S.H.E.
Abstract:Matrix-assisted laser desorption/ionization-mass spectrometry peptide mass mapping and nano-electrospray ionization tandem mass spectrometry were used to identify acidic, low molecular mass proteins of Mycobacterium tuberculosis strain H37Rv. Proteins were extracted from whole cell lysates of mycobacteria, separated by high resolution two-dimensional electrophoresis (2-DE) and analysed by mass spectrometry (MS). Silver-stained 2-DE patterns resolved about 1800 distinct protein species, 190 of which had an observed isoelectric point and molecular mass in the range of pH 4 to 6 and 6 to 15 kDa, respectively. Seventy-six spots from this range were excised from Coomassie Brilliant Blue G250-stained gels and analysed by MS, from which 72 were identified. These spots were shown to represent products of as many as 50 different protein-coding genes. Ten genes gave rise to more than one protein species. Eleven spots contained more than one protein. The present study led to the identification of 15 mycobacterial proteins with assigned putative functions, 28 conserved hypothetical proteins and one unknown protein. Most proteins of the latter two groups had previously been predicted at the DNA level only. Six additional spots were shown to comprise proteins encoded by open reading frames that have not been predicted for M. tuberculosis H37Rv by genomic investigations.
Keywords:Low molecular Mass Proteins, Mass Spectrometry, Mycobacterium Tuberculosis, Peptide Mass Mapping, Proteomics, Two-Dimensional Electrophoresis
Source:Proteomics
ISSN:1615-9853
Publisher:Wiley
Volume:1
Number:4
Page Range:494-507
Date:1 January 2001
Official Publication:https://doi.org/10.1002/1615-9861(200104)1:4<494::AID-PROT494>3.0.CO;2-4
PubMed:View item in PubMed

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