<i>Mycobacterium tuberculosis</i>in the Proteomics Era

Gengenbacher, Martin; Mouritsen, Jeppe; Schubert, Olga T.; Aebersold, Ruedi; Kaufmann, Stefan H. E.

doi:10.1128/microbiolspec.mgm2-0020-2013

Cited by 18 publications

(20 citation statements)

References 161 publications

(166 reference statements)

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“…This process is performed most commonly by spectral mass-matching, and occasionally by de novo sequencing [73]. The bottom-up approach has been used as a building block for both discovery and targeted proteomics and widely applied to basic mycobacterial research [52,74,75]. These discovery-based methods have aided in profiling clinical isolates of M. tuberculosis and distinguishing the relationship between protein synthesis and active disease [19,[76][77][78].…”

Section: Targeted and Comprehensive Proteomic Approaches Define The Mmentioning

confidence: 99%

The genetic proteome: Using genetics to inform the proteome of mycobacterial pathogens

et al. 2021

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Mycobacterial pathogens pose a sustained threat to human health. There is a critical need for new diagnostics, therapeutics, and vaccines targeting both tuberculous and nontuberculous mycobacterial species. Understanding the basic mechanisms used by diverse mycobacterial species to cause disease will facilitate efforts to design new approaches toward detection, treatment, and prevention of mycobacterial disease. Molecular, genetic, and biochemical approaches have been widely employed to define fundamental aspects of mycobacterial physiology and virulence. The recent expansion of genetic tools in mycobacteria has further increased the accessibility of forward genetic approaches. Proteomics has also emerged as a powerful approach to further our understanding of diverse mycobacterial species. Detection of large numbers of proteins and their modifications from complex mixtures of mycobacterial proteins is now routine, with efforts of quantification of these datasets becoming more robust. In this review, we discuss the “genetic proteome,” how the power of genetics, molecular biology, and biochemistry informs and amplifies the quality of subsequent analytical approaches and maximizes the potential of hypothesis-driven mycobacterial research. Published proteomics datasets can be used for hypothesis generation and effective post hoc supplementation to experimental data. Overall, we highlight how the integration of proteomics, genetic, molecular, and biochemical approaches can be employed successfully to define fundamental aspects of mycobacterial pathobiology.

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Section: Targeted and Comprehensive Proteomic Approaches Define The Mmentioning

confidence: 99%

The genetic proteome: Using genetics to inform the proteome of mycobacterial pathogens

et al. 2021

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“…Supplementary Figure 1: Spot numbers of protein spots selected after 2D-DIGE analysis and identified by MALDI-TOF/TOF MS. GarA and GlnA1 spots were previously identified (spots numbers 1,3,4,12,15,19,21,25,30,35,37,40) (24).…”

Section: Supplementary Figurementioning

confidence: 99%

Proteome remodeling in theMycobacterium tuberculosisPknG knockout: molecular evidence for the role of this kinase in cell envelope biogenesis and hypoxia response

Lima

Leyva

Rivera

et al. 2021

Preprint

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Mycobacterium tuberculosis, the ethiological agent of tuberculosis, is among the deadliest human pathogens. One of M. tuberculosis pathogenic hallmarks is its ability to persist in a dormant state in the host for long periods, reinitiating the infectious cycle when favorable environmental conditions are found. Thus, it is not surprising that this pathogen has developed different mechanisms to withstand the stressful conditions found in the host. In particular, the Ser/Thr protein kinase PknG has gained special relevance since it regulates nitrogen metabolism and facilitates bacterial survival inside macrophages. Nevertheless, the molecular mechanisms underlying these effects are far from being elucidated. To further investigate these issues, we performed quantitative proteomics analyses of protein extracts from M. tuberculosis H37Rv and a mutant derivative lacking pknG. Our results showed that in the absence of PknG the mycobacterial proteome was remodeled since 5.7% of the proteins encoded by M. tuberculosis presented significant changes in its relative abundance when compared to the wild-type strain. The main biological processes affected by pknG deletion were the biosynthesis of cell envelope components and the response to hypoxic conditions. As many as 13 DosR-regulated proteins were underrepresented in the pknG deletion mutant, including the distinctive Hrp-1, which was found to be 12-fold decreased according to Parallel Reaction Monitoring experiments. Altogether, the results presented here allow us to postulate that PknG regulation of bacterial adaptation to stress conditions might be an important mechanism underlying its reported effect on intracellular bacterial survival.

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“…Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)using separated proteins by one-or two-dimensional polyacrylamide gel electrophoresis (PAGE)has been used extensively to identify, quantify and compare isolates showing different phenotypes [6][7].MALDI-TOF-MS has become a widely used technique for analysis of proteins and peptides due to its operational simplicity, robustness and high sensitivity and atypical analysis consists of i-separation of protein(s)in a mixture by PAGE, iivisualization of separated protein(s), iii-excision and in-gel digestion of protein(s), iv-analysis of the proteolytically digested peptides by MALDI-MS resulting in a map of peptide mass [7]. The use of this technique has provided the means for comparison of differentially expressed bacterial proteins under various culture conditions and by strains with different genotypes, thus supplying a more comprehensive view of the life of a pathogen [8].However, despite the advances in the field of MTB proteomics there are few reported literature dealing with differential expression of proteins in multidrug resistant bacteria compared to susceptible cells [9].In this study protein expression profile of MDR and sensitive isolates of MTB were analyzed and compared in order to identify proteins, which could be used in prevention, diagnosis and treatment.…”

Section: Introductionmentioning

confidence: 99%

Protein profiling and analysis of drug sensitive and multidrug resistant isolates of Mycobacterium tuberculosis by native polyacrylamide gel electrophoresis and mass spectrometry

Yari

Tasbiti

Ghanei

et al. 2015

vacres

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Introduction: Tuberculosis (TB) remains a deadly infectious disease despite all the efforts to reduce its incidence. Spread of multidrug resistant TB has seriously undermined the efforts to control the disease globally. In this study protein expression profile of MDR and sensitive isolates of MTB were analyzed and compared in order to identify proteins, which could be used in prevention, diagnosis and treatment. Methods: A sensitive and MDR isolate of Mycobacterium tuberculosis (MTB) were cultured on Middlebrook 7H9 medium and the whole cell lysates were subjected to native polyacrylamide gel electrophoresis (NPAGE) for protein expression profiling. Protein bands present in the MDR cell lysate that were not detected in the sensitive cell lysate were sent for identification by Matrixassisted laser desorption/ionization time-of-flightmass spectrometry (MALDI-TOF-MS). Results: Comparison of the protein expression profiles showed 6 bands that were not detected in the sensitive isolates. MTB Structural Annotation database search of the mass spectrometry results identified these bands as Rv3597c, Rv0379,Rv3614c, Rv0475, Rv0462, andRv0147and global transcriptional regulation, involvement in cell wall and cell processes and intermediary metabolism and respiration were the functions attributed to these proteins. Conclusion: Our results highlighted the complexities of linking protein expression to MDR phenotype as none of the proteins identified could be linked directly to drug resistance. The proteins identified in the present study were mostly those essential for survival or virulence of the bacteria, and could be used for diagnosis or as candidate vaccine, but with a better understanding of the function of these proteins their association with the MTB resistance to antibiotics might become clear.

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Mycobacterium tuberculosisin the Proteomics Era

Cited by 18 publications

References 161 publications

The genetic proteome: Using genetics to inform the proteome of mycobacterial pathogens

The genetic proteome: Using genetics to inform the proteome of mycobacterial pathogens

Proteome remodeling in theMycobacterium tuberculosisPknG knockout: molecular evidence for the role of this kinase in cell envelope biogenesis and hypoxia response

Protein profiling and analysis of drug sensitive and multidrug resistant isolates of Mycobacterium tuberculosis by native polyacrylamide gel electrophoresis and mass spectrometry

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