Comparative proteome analysis of Mycobacterium tuberculosis and Mycobacterium bovis BCG strains: towards functional genomics of microbial pathogens

Abstract

SummaryIn 1993, the WHO declared tuberculosis a global emergency on the basis that there are 8 million new cases per year. The complete genome of the strain H37Rv of the causative microorganism, Mycobacterium tuberculosis, comprising 3924 genes has been sequenced. We compared the proteomes of two non-virulent vaccine strains of M. bovis BCG (Chicago and Copenhagen) with two virulent strains of M. tuberculosis (H37Rv and Erdman) to identify protein candidates of value for the development of vaccines, diagnostics and therapeutics. The mycobacterial strains were analysed by two-dimensional electrophoresis (2-DE) combining non-equilibrium pH gradient electrophoresis (NEPHGE) with SDS±PAGE. Distinct and characteristic proteins were identi®ed by mass spectrometry and introduced into a dynamic 2-DE database (http:/ /www.mpiibberlin.mpg.de/2D-PAGE). Silver-stained 2-DE patterns of mycobacterial cell proteins or culture supernatants contained 1800 or 800 spots, respectively, from which 263 were identi®ed. Of these, 54 belong to the culture supernatant. Sixteen and 25 proteins differing in intensity or position between M. tuberculosis H37Rv and Erdman, and H37Rv and M. bovis BCG Chicago, respectively, were identi®ed and categorized into protein classes. It is to be hoped that the availability of the mycobacterial proteome will facilitate the design of novel measures for prevention and therapy of one of the great health threats, tuberculosis.

Liberation of Soluble Proteins from Live and Dead Mycobacterial Cells and the Implications for Pathogenicity of Tubercle Bacilli

“…Figure 2 illustrates the distribution of proteins in live M. tuberculosis. The two-dimensional polyacrylamide gel electrophoresis patterns of his cultures are therefore relatively simple with few protein species compared to those of for example Jungblut [13]. The only comparable culture filtrates known, which probably had the same quality to those of Sadamu Nagai, were those of Fukui et al [18].…”

Comparative proteome analysis of Mycobacterium tuberculosis grown under aerobic and anaerobic conditions

“…Furthermore, comparative proteome analysis of M. tuberculosis and M. bovis BCG has shown differences in the expression of several protein classes (Jungblut et al, 1999). There are also differences in protein expression between different laboratory strains, such as M. tuberculosis Erdman and H37Rv (Jungblut et al, 1999).…”

“…There are also differences in protein expression between different laboratory strains, such as M. tuberculosis Erdman and H37Rv (Jungblut et al, 1999). To study latency it is therefore important to use a model with a relevant experimental M. tuberculosis strain.…”

Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry

“…The newly identified antigens are being used to develop vaccines, e.g. for pathogenic bacteria, such as H. pylori and M. tuberculosis [121][122][123][124]. New data on cell differentiation and on cell regulation can be obtained using the total cell protein approach, and new insights in functional genomics can be derived using these data.…”

“…For the elucidation of immunologically relevant proteins, the proteomes of M. tuberculosis and Heliobacter pylori were studied by high-resolution 2-DE and MALDI mass fingerprinting of the virulent and the attenuated strains [121][122][123]. From the mycobacterial unique spots, 36 were evaluated in a mouse model as DNA vaccines and one vaccine improved the efficacy of BCG vaccination [124].…”