What Can Proteomics Tell Us about Tuberculosis?
What Can Proteomics Tell Us about Tuberculosis?
Journal of Microbiology and Biotechnology. 2015. Sep, 25(8): 1181-1194
Copyright © 2015, The Korean Society For Microbiology And Biotechnology
  • Received : February 04, 2015
  • Accepted : February 26, 2015
  • Published : September 28, 2015
Export by style
Cited by
About the Authors
Susana, Flores-Villalva
INIFAP, Centro Nacional de Investigación Disciplinaria en Fisiología y Mejoramiento Animal, Colón, Qro, México
Elba, Rogríguez-Hernández
INIFAP, Centro Nacional de Investigación Disciplinaria en Fisiología y Mejoramiento Animal, Colón, Qro, México
Yesenia, Rubio-Venegas
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Querétaro, Qro, México
Jorge Germinal, Cantó-Alarcón
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Querétaro, Qro, México
Feliciano, Milián-Suazo
Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Querétaro, Qro, México

Tuberculosis (TB) is an infectious disease transmitted by aerosol droplets and characterized by forming granulomatous lesions. Although the number of people infected in the population is high, the vast majority does not exhibit symptoms of active disease and only 5-10% develop the disease after a latent period that can vary from weeks to years. The bases of the immune response for this resistance are unknown, but it depends on a complex interaction between the environment, the agent, and the host. The analysis of cellular components of M. tuberculosis shows important host-pathogen interactions, metabolic pathways, virulence mechanisms, and mechanisms of adaptation to the environment. However, the M. tuberculosis proteome still remains largely uncharacterized in terms of virulence and pathogenesis. Here, we summarize some of the major proteomic studies performed to scrutinize all the mycobacterial components.
Tuberculosis (TB) is the second leading cause of death from a single infectious agent, after the immunodeficiency virus (HIV). In 2013, 9.0 million people developed the disease and 1.5 million died, and 25% of them were HIV positive [106] . In fact, TB is the leading killer of people infected with HIV, causing one fifth of all deaths in this population. Most deaths due to TB occur in men; however, TB is the third most important cause of death in women; half of the deaths in the HIV-positive group are woman. Six percent of all cases are children under 15, and 8% of them die. Currently, multidrug-resistant TB is present in almost all countries surveyed [106] .
Tuberculosis is caused by Mycobacterium tuberculosis , a member of the M. tuberculosis complex, which also includes M. bovis, M. microti, M. caprae, M. canettii, and M. africanum [26] , most of them capable of causing disease in humans. Typically, TB affects the lungs and the lymph nodes, but it can actually affect any organ in the body [53] . Infection establishes in approximately one third of the individuals exposed to the tuberculous bacillus; however, only about 10% ever develop the disease [16 , 45 , 76] .
There are quite a few reviews in the literature about studies analyzing the complex immune response towards M. tuberculosis in infected individuals, but only a few explore the interaction between the host cells and the bacilli [25 , 68 , 74 , 81 , 85] . There are also studies that review the most important mycobacterial components during infection [29 , 33 , 34 , 37] . However, the proteome of M. tuberculosis remains unclear, especially in terms of virulence and pathogenesis [56] , in spite of the increasing interest in detecting mycobacterial antigens related to the immune responses in skin testing. Before the 90’s, many mycobacterial proteins were identified using methods based essentially on biochemical fractionation and on immunological screening with monoclonal antibodies in patient sera. At that time, important proteins such as the Ag85 complex, MPB64, MPB70, and some cytoplasmic proteins like DnaK, GroEl, and SodA were identified [18 , 112] .
In the last two decades, there has been increasing interest in studying the proteome of M. tuberculosis . Classical studies involve two-dimensional electrophoresis (2-DE), in which proteins are separated according to charge and molecular weight. This method resolves typically between 1,500 and 3,000 spots [105] . Protein spots can be visualized with silver or Coomassie staining or by fluorescent dyes. Every spot is then isolated and digested with trypsin to produce 10 mer peptides, which are then subjected to techniques like mass spectrometry (MS), tandem MS (MS/MS); matrix-assisted laser desorption/ionization mass spectrometry (MALDI/MS), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Nagai et al . [67] were able to purify and partially characterize 12 different proteins from culture filtrate of M. tuberculosis using one- and two-dimensional electrophoresis to monitor the separation of individual proteins during various chromatographic procedures. Two-dimensional liquid chromatography (LC-MS/MS) and the isotope-coded affinity tag technology (ICAT) have also been reported in proteomic studies [10 , 40] . Other authors combined 2-DE, western blotting, N-terminal amino acid sequencing, and liquid chromatography-mass spectrometry (LC-MS-MS) to identify 205 spots and map 32 N-terminal sequences [96] . These original studies had led to the creation of 2-DE maps, but it was until the description of the complete genome sequence of M. tuberculosis that proteomic analysis really helped to understand this pathogen. In this review we summarize the state of art about the proteomics of M. tuberculosis .
Cellular Components
The analysis of cellular components of M. tuberculosis shows important host-pathogen interactions, metabolic pathways, virulence mechanisms, and mechanisms of adaptation to the environment. For example, the analysis of culture filtrates allows the identification of proteins secreted by the bacilli, potential markers of virulence, or immunogenic antigens for the development of new vaccines and diagnostic tools. Cell wall and membrane proteins are involved in the resistance of the pathogen to chemical injury and play a fundamental role in hostpathogen interactions; therefore, the study of the cell wall proteins can be useful in the development of new drugs. Some of the proteins identified in the studies described below are listed in Table 1 .
Description of some proteins identified in the studies described in this review.
PPT Slide
Lager Image
aIt is possible that some proteins described by other authors are not listed here.
Membrane Proteins
Initial interactions between M. tuberculosis and the host mark the pathway of infection and the subsequent immune host-response that defines disease outcome. These interactions include many important membrane surface enzyme/transporters involved in intercellular multiplication and bacterial response to host microbicidal processes [17] . Membrane proteins are active in cell-to-cell interactions, ion transport, and cell signaling. Thus, more than half of all membrane proteins are predicted to be pharmaceutical targets [86] . Many proteins related to M. tuberculosis pathogenicity are surface proteins involved in lipid metabolism and transport across the membrane [34] . Identification of these proteins can help to better understand the bacterial mechanisms of virulence and help in the discovery of new drug targets. For example, some lipoproteins and glycolipoproteins (19 kDa protein, LprG, PstS1) interact with TLR2 receptors and modulate host immune responses affecting antigen presentation function of macrophages [38 , 39] . In order to gain access through different tissues, bacteria have evolved a wide range of molecules to enable them to bind to select host molecules like proteoglycans, fibronectin, and laminin. Some of these molecules are the proteins of antigen 85 complex, Apa, Wag22, HbhA, and others [25] ( Table 1 ).
Unfortunately, membrane protein characterization is difficult to accomplish, as the proteins are molecules composed of both hydrophobic and hydrophilic regions, so no single solvent or combination of solvents can solubilize all membrane proteins. Two-dimensional technology can underestimate the number of membrane proteins and membrane-associated proteins because of their low solubility. For that reason, various authors have changed the traditional 2-DE approach with 1-D SDS-PAGE technology. Gu et al . [43] were able to identify 739 membrane and membrane-associated proteins. They first obtained the membrane subcellular fraction of M. tuberculosis through a differential centrifugation method. Then, they separated this fraction on a 1-D SDS gel and treated each gel slide with trypsin, and then the peptides digested were analyzed by high-resolution microcapillary LC separation prior to a highly sensitive nanospray-MS/MS analysis. Very hydrophobic proteins, even those with 15 trans-membrane helices, were detected with this methodology.
Studies of membrane proteins have shown that the plasma membrane is easily contaminated by cytosolic and secreted proteins, which could be part of protein complexes. Xiong et al . [108] proposed a method to remove cytosolic proteins. It consisted of treating the plasma membrane with 5 M urea and a high pH carbonate solution, followed by ultracentrifugation. With this method, membrane proteins, together with the phospholipid bilayer, aggregate to form a pellet, whereas proteins weakly associated or bound to plasma membrane proteins and contaminants remain in the supernatant. By following this method, coupled with 1-DSDS-PAGE and nanoLC electrospray ionization MS/MS, it has been possible to identify 349 proteins, 100 of which were integral membrane proteins with at least one predicted transmembrane α-helix.
Triton X-114 has also been used to remove lipids or carbohydrates from the membrane. These components could interfere with the protein resolution [94] . The Triton X-114 method, in combination with nano-ESI-LC-MS/MS for protein extraction, allowed the identification of 43 predicted lipoproteins [58] . It has been suggested that lipoproteins are involved in host-pathogen interactions [97] . These lipoproteins have also been identified as strong immunoantigens for serodiagnosis of tuberculosis [107] . With this methodology, the 10 most abundant lipoproteins in M. tuberculosis membranes have been identified, and only three (Rv0432, Rv3763, and Rv0932c) have shown some biological function. These authors also describe one major lipoprotein of M. tuberculosis , Rv3623, which has become a pseudogene in M. bovis [59] .
Shotgun proteomics, labeled and label free, provide a new dimension to proteomic methods [71] . It helps in identifying differences in protein composition between attenuated and virulent strains. In order to compare the membrane protein expression profiles of M. tuberculosis H37Rv and its attenuated counterpart H37Ra, Målen et al . [60] used orbitrap mass spectrometry technology in combination with relative protein expression abundance calculations. A total of 1,771 different protein groups were identified, of which 1,300 were common to both strains, 278 were exclusively identified in M. tuberculosis H37Rv, and 193 were only observed in the H37Ra strain. Although the majority of the identified proteins had similar relative abundance in both strains, 19 membrane and lipoproteins had higher abundance in H37Rv.
Cell Lysates and Cytoplasmic Proteins
After the whole sequence of the M. tuberculosis genome, reference maps of cellular fractions have been reported [82] . A subcellular fractionating protocol, which includes sonication, mechanical disruption, and differential centrifugation, was applied to generate 2-DE maps of the cell wall, cytosol proteins, and culture filtrates. In total, 1,184 spots from the three fractions were observed; the protein profile of the three elements was quite different. Sixty-one proteins were identified, and 31 were novel proteins. A genomics non-predicted protein was found, TB9.4, and revealed the presence of an alternative star codon [84] . Another set of six genomics non-predicted proteins have been identified, showing how genomics and proteomics complement each other [49] . Using a different procedure, subcellular fractionation in combination with membrane enrichment using wheat germ agglutinin (WGA) and LC-MS/MS analysis, 1,051 proteins of M. tuberculosis were identified, including 183 transmembrane proteins [4] .
Because of the complexity of the proteome, proteome-wide studies are still limited in their ability to quantify and identify all proteins. However, recently, the identification of 3,434 proteins from M. bovis BCG has been reported, including 512 transmembrane proteins, which represent about 87% of the proteins expressed in the BCG strain. In this study, three subcellular fractions were separated: cell wall, plasma membrane, and cytoplasm. The proteins extracted for each fraction were resolved by 1-D SDS-PAGE. Then, each lane was cut and digested, and peptides were separated by reversed-phase chromatography and analyzed using a LTQ Orbitrap Velos. Up to now, this is the most comprehensive study of the BCG proteome; it will help in the design of vaccination and immunodiagnostic strategies against TB [113] .
Culture Filtrate Proteins
Analysis of secreted proteins in culture filtrates has been difficult owing to the tendency of the mycobacterial culture to undergo autolysis with the subsequent release of cytoplasmic proteins to the culture filtrate [82 , 103] . However, it was possible to identify some of the major M. tuberculosis secreted proteins with the use of a strain that did not undergo the normal autolytic process; the culture filtrates were therefore enriched with secreted proteins [36] . Secreted proteins have been suggested as host protective antigens responsible for the rapid recognition of the bacilli by host lymphocytes. These proteins are available to the immune system more easily when the vaccine used is alive than when it is dead [46] . The most abundant proteins found up to now in culture filtrates have been antigen 85 complex (Ag85A, Ag85B, Ag85C, and MPT51 or Ag85D), MPT63, MPT64, MPT32, MTC28, ESAT-6, CFP10, and others members of the ESAT-6 family [57] .
Recently, 103 novel BCG secreted proteins have been reported. The identification of these proteins was accomplished through the use of one-dimensional gel electrophoresis and high-resolution tandem mass spectrometry. These proteins represented potential predominant antigens in humoral and cellular immune responses [115]. Deenadayalan et al . [22] identified two new T cell antigens (AcpM and PpiA) through the fractioning of culture filtrates by preparative isoelectric focusing and the analysis of each fraction by immunological studies.
Comparative Proteomics
Proteins present in M. tuberculosis but absent in M. bovis BCG are valuable antigens for novel diagnostic, therapeutic, and vaccination strategies. Because of this, comparative proteomic analysis focuses on differentiating the protein composition between virulent and attenuated mycobacterial strains. Jungblut et al . [50] compared proteins present in two virulent laboratory M. tuberculosis strains (H37Rv and Erdman) with those present in two M. bovis BCG strains (Chicago and Copenhagen) using 2-DE and MALDI/MS analysis. As expected, the protein composition was very similar, but there were clear differences in spot intensity, and presence or absence and position of the spots. In comparison with BCG, H37Rv comprised 13 additional spots and lacked eight spots. Nine spots had lower intensity, and one spot had higher intensity. Six spots were unique in H37Rv: L -alanine dehydrogenase (Rv2780), isopropyl malate synthase (Rv3710), nicotinate-nucleotide pyrophosphatase (Rv1596), MPT64 (Rv1980c), and two conserved hypothetical proteins (Rv2449c, Rv0036c). Eight proteins were unique to BCG.
Subsequently, using 2-DE and MS, 56 unique proteins were detected in M. tuberculosis and 40 in M. bovis BCG. Twelve M. tuberculosis -specific proteins were identified as products of genes reported to be missing in BCG [60] . The same group compared culture supernatant proteins from M. tuberculosis H37Rv and M. bovis BCG, and 27 M. tuberculosis -specific proteins and 22 novel differential proteins, such as acetyl-CoA C-acetyltranferase and two putative ESAT-6 like proteins, were described [63] .
Proteomics has been used to understand differences in protection of the different substrains of BCG. This has been done by comparing the membrane protein fractions of M. tuberculosis and M. bovis BCG. The authors were able to detect changes in LpqH, Icl1, and GlcB proteins between these strains. Besides this, it was showed that levels of ESX-3 in BCG were reduced compared with M. tuberculosis ; and because the ESX-3 system is essential in M. tuberculosis with an apparent function in iron and zinc homeostasis, it raises the possibility that the vaccination efficacy of BCG could be improved by increasing the levels of the ESX-3 system [44] .
Proteomic analysis of M. tuberculosis strains with differences in virulence or drug resistance may provide clues in relation to pathogenicity and help in the identification of virulence factors. Using MS and label-free quantitative proteomic approaches to estimate differences in protein abundance in two clinical Beijing strains, it was found that 48 proteins were over-represented in the hypovirulent isolates, and 53 were over-represented in the hypervirulent. These isolates displayed differences in their level of virulence as defined by their epidemiological and population characteristics as well as virulence in a mouse model of infection. Molecules of cell wall organization and DNA transcription regulatory proteins may have a critical influence in defining the level of virulence [20] .
In an effort to determine protein differences between isoniazid resistant and susceptible M. tuberculosis strains, Jiang et al . [47] compared the proteins extracted from nine isoniazid monoresistant and seven isoniazid susceptible M. tuberculosis strains. Five proteins that were up-regulated in isoniazid-resistant strains were characterized by MS: Rv1446c, Rv3028c, Rv0491, Rv2145c, and Rv2971. Most of these differentially expressed proteins are membrane proteins. The gene Rv0491 encodes the protein RegX3, which belong to SenX3-RegX3 of the two-component regulatory system. These systems enable organisms to respond to changing environmental conditions and are involved in M. tuberculosis virulence [47 , 60] .
Latent-Associated Proteins
M. tuberculosis is one of the most successful pathogens of humans. This success is linked to its ability to persist within the human body for long periods of time without causing any symptoms of disease, in a state of latency. The actual state of the bacterium in latency is not well understood; however, different models in vitro have been developed in order to mimic the in vivo conditions during latent infection. Conditions such as nutrient restriction, low pH, and reduced oxygen tensions have been used. Identification of proteins expressed during latency is important to study the mechanisms of this state. These proteins could represent important targets for vaccine and drug development [11] .
The Wayne model of dormancy was used to analyze BCG proteins induced in hypoxic stationary phase cultures. 2-DE analysis showed an up-regulation of the HspX protein, as well as the DosR regulator (Rv3133c) [8] . Metabolic labeling of BCG showed that at least 45 proteins were differentially expressed under standing and shaking culture conditions; the greatest induction was observed with Rv2623, which was expressed under standing conditions but not in shaking cultures [31] . It is interesting that Rv2623 has the ability to regulate growth in vitro and in vivo , and that it is necessary for the establishment of a persistent infection [23] . Comparative analysis of protein content between aerobic and anaerobic cultures of M. tuberculosis showed 13 unique proteins and 37 proteins that were more abundant during anaerobic conditions, some of them related to mycolic acid synthesis and oxidative stress [99] . Two additional proteins (Fba (Rv0363c) and Ald (Rv2780)) were found in large amounts in culture filtrates of M. tuberculosis grown under oxygen limitation. These proteins are implicated in glycolysis and cell wall synthesis [83] . Cho et al . [13] compared the protein profile of the NRP-1 and NRP-2 stages of metabolic adaptation induced in the Wayne model of hypoxia, applying ICAT technology. They were able to identify 586 proteins in the microaerobic stage NRP-1, and 628 proteins in the anaerobic stage NRP-2. The analysis showed that protein expression decreased in NRP-1 relative to NRP-2, suggesting a temporal down-regulation of the translation machinery necessary for adaptation to low oxygen concentrations.
Through label-free LC MS/MS analysis and applying a nutrient starvation model, 1,176 proteins were identified in culture filtrates of M. tuberculosis . The levels of 230 proteins increased in nutrient-starved culture, and the levels of 208 proteins decreased. In this study, proteins of the toxinantitoxin systems were present in large quantities in nutrient-starved cultures, supporting the hypothesis of its role as metabolic switches [2] . Finally, a comprehensive proteomic analysis of M. tuberculosis over the mid-log, early stationary, and late stationary phases was performed. Ten proteins were differentially expressed in the late stationary phase compared with the other two phases, which involve proteins belonging to metabolic pathways and cell wall or lipid biosynthesis [3] .
Intracellular Proteins ofMycobacterium
M. tuberculosis is an intracellular pathogen that has developed mechanisms to resist the hostile environment inside the macrophage. This resistance involves genes that are potential virulence factors, as well as important proteins for interaction with the host cell. More than that, intracellular mycobacterial proteins can be processed and presented by infected macrophages, so they can represent important protective immune factors; nevertheless, there are only a few studies of mycobacterial proteins expressed in vivo , in spite of the valuable information they could provide about natural infection.
The studies of intracellular proteins of mycobacteria are limited because of the difficulties for recovering enough amounts of mycobacterial proteins free from contaminating host cells. Using metabolic labeling of M. tuberculosis and infection of THP-1 cells, Lee and Horwitz [55] identified 44 proteins expressed differentially, where 16 were up-regulated and 28 down-regulated. Of the 16 up-regulated proteins, six were absent during extracellular growth under normal and stress conditions. In a different study, six abundant proteins showed increased expression inside the macrophage: 16 kDa α-crystallin (HspX), GroEL1, GroEL2, Rv2623, InhA, and elongation factor Tu (Tuf). In this study, it was also shown that most of the intracellular modulation of protein expression occurred within the first 12-24 h following phagocytosis, reflecting the dynamic intracellular environment during the interaction between the cell and the bacilli [66] . Using subcellular fractionation of infected macrophages to isolate phagosomal M. tuberculosis , in combination with high-resolution 2-DE and MALDI-MS, 11 exclusively intraphagosomal mycobacterial proteins were detected. Some were involved in metabolism and cell envelope synthesis [64] .
In order to understand whether the mechanisms adopted by resistant and sensitive mycobacteria to survive and grow inside the macrophage are similar or different, Singhal et al . [93] analyzed the protein profile of M. tuberculosis MDR and sensitive clinical isolates infecting THP-1 cells. Mass spectrometry and bioinformatic characterization showed that the majority of commonly expressed proteins belonged to the cellular metabolism and respiration category. More than that, some common proteins were found to be overexpressed, so it is possible that some common mechanism is adopted by sensitive and resistant strains for their survival inside the macrophage. Understanding this mechanism could be useful in drug targeting [93] .
Recently, the first proteomic characterization of M. tuberculosis during infection in vivo in a guinea pig model by aerosol infection was reported, and over 500 proteins present at 30 and 90 days post infection were described. Two functional groups represent about half of the total proteins identified: category 3 (cell wall and cell process) and category 7 (intermediary metabolism and respiration). These proteins displayed high heterogeneity, indicating important biological processes necessary in different stages of infection. Many proteins identified in this study may provide the basis for rational drug design [52] .
PPD Protein Composition
Tuberculin skin testing (TST) is the gold standard for determining whether an individual is infected with TB. This test involves the intradermal injection of a purified protein derivative (PPD), which is a poorly defined mix of proteins, and little is known regarding which of these components are responsible for the DTH response. A complete knowledge of the molecular composition of the PPD will allow the selection of proteins specific for a more refined test to human and bovine TB. However, little has been done to identify the components of the PPD, because of the degradation, partial denaturation, and aggregation of many of the protein components [110] .
Borsuk et al . [9] characterized the proteins present in bovine and avium PPD from Brazil and UK. A total of 171 different proteins were identified, most of them cytoplasmic proteins related to intermediary metabolism and respiration. Most secreted proteins were present in the PPD preparation: Ag85 complex, MPT32, MPT64, MPT83, MPT53, and MPT70. Some differences between PPD preparations from Brazil and UK were observed, probably due to differences in preparation methods and these partially could explain the differences in biological potency of PPD products from different sources [110] . Twenty-one proteins were identified in both bovine PPD preparations, but not in M. avium preparations, where 10 of these were not present in M. avium ; therefore, these proteins are candidates to differentiate exposure of the animal to environmental mycobacteria from bovine tuberculosis [9] .
A proteomic analysis of the FDA standard PPD-S2 showed that it is composed of at least 240 proteins, and many of the known M. tuberculosis T cell antigens dominated the PPD composition (GroES, GroEL2, HspX, and DnaK). Additional comparative proteomic and histological analysis between PPD-S2, RT23, and PPD-KIT demonstrated that differences in the relative abundance of several proteins, including members of the Esx protein family, may contribute to the increased inflammatory responses observed with RT23 and PPD-KIT reagents, supporting the theory that variability in PPD reagents may explain the differences in DTH responses reported among populations [14] . Protein analysis of PPD-CT68 (Tubersol) showed that 142 proteins were shared between PPD-CT68 and PPD-S2 preparations, whereas 123 and 89 were exclusively identified from PPD-CT68 and PPD-S2, respectively. Eighteen proteins were common in PPDs from M. tuberculosis, M. bovis, and M. avium [75] .
Despite the identification of many antigens, it is still challenging to replace classical PPD preparations. Currently, ESAT-6, CFP-10, Rv3615, DnaK, and GroEL2 are under evaluation as next-generation PPD candidates in the diagnosis of human [6 , 111] and bovine TB [32 , 91 , 101] .
M. tuberculosis is a very successful pathogen that has a very complex interaction with the host cell, where it can persist for long periods without causing any symptoms of disease. WHO estimates that almost one-third of the world population is latently infected with M. tuberculosis and this latency is a constant source of disease reactivation. Five to ten percent of the infected individuals develop active TB over their lifetime, and defects in cell-mediated immunity, HIV co-infection, malnutrition, administration of chemotherapy or antitumor necrosis factor therapy, and diabetes predispose latently infected people to develop TB [16] . Therefore, it is quite important to develop new diagnostic reagents, new vaccines, and new targets for TB drugs. Proteomics can have a significant impact on our current understanding of M. tuberculosis , especially about its pathogenicity, virulence, and interactions with host cells.
This work was partially supported by FORDECYT No. 193512. We gratefully acknowledge the revision of the English language usage performed by Eline Boele.
Abdallah AM , Gey van Pittius NC , Champion P a D , Cox J , Luirink J , Vandenbroucke-Grauls CMJE 2007 Type VII secretion--mycobacteria show the way. Nat. Rev. Microbiol. 5 883 - 891    DOI : 10.1038/nrmicro1773
Albrethsen J , Agner J , Piersma SR , Højrup P , Pham T V , Weldingh K 2013 Proteomic profiling ofMycobacterium tuberculosisidentifies nutrient-starvation-responsive toxinantitoxin systems. Mol. Cell. Proteomics 12 1180 - 1191    DOI : 10.1074/mcp.M112.018846
Ang K , Ibrahim P , Gam L 2014 Analysis of differentially expressed proteins in late-stationary growth phase ofMycobacterium tuberculosisH37Rv. Biotechnol. Appl. Biochem. 61 153 - 164    DOI : 10.1002/bab.1137
Bell C , Smith T , Sweredoski MJ , Hess S 2012 Characterization of theMycobacterium tuberculosisproteome by liquid chromatography mass spectrometry-based proteomics techniques: a comprehensive resource for tuberculosis research. J. Proteome Res. 11 119 - 130    DOI : 10.1021/pr2007939
Berger BJ , Knodel MH 2003 Characterisation of methionine adenosyltransferase fromMycobacterium smegmatisandM. tuberculosis. BMC Microbiol. 3 1 - 13    DOI : 10.1186/1471-2180-3-12
Bergstedt W , Tingskov PN , Thierry-Carstensen B , Hoff ST , Aggerbeck H , Thomsen VO 2010 First-in-man open clinical trial of a combined rdESAT-6 and rCFP-10 tuberculosis specific skin test reagent. PLoS One 5 e11277 -    DOI : 10.1371/journal.pone.0011277
Bhatt A , Fujiwara N , Bhatt K , Gurcha SS , Kremer L , Chen B 2007 Deletion of kasB inMycobacterium tuberculosiscauses loss of acid-fastness and subclinical latent tuberculosis in immunocompetent mice. Proc. Natl. Acad. Sci. USA 104 5157 - 5162    DOI : 10.1073/pnas.0608654104
Boon C , Li R , Qi R , Dick T 2001 Proteins ofMycobacterium bovisBCG induced in the wayne dormancy model. J. Bacteriol. 183 2672 - 2676    DOI : 10.1128/JB.183.8.2672-2676.2001
Borsuk S , Newcombe J , Mendum TA , Dellagostin OA , McFadden J 2009 Identification of proteins from tuberculin purified protein derivative (PPD) by LC-MS/MS. Tuberculosis (Edinb.) 89 423 - 430    DOI : 10.1016/
Brewis IA , Brennan P 2010 Proteomics technologies for the global identification and quantification of proteins. Adv. Protein Chem. Struct. Biol. 80 1 - 44
Chao MC , Rubin EJ 2010 Letting sleepingdoslie: does dormancy play a role in tuberculosis? Annu. Rev. Microbiol. 64 293 - 311    DOI : 10.1146/annurev.micro.112408.134043
Chen JM , Boy-Röttger S , Dhar N , Sweeney N , Buxton RS , Pojer F 2012 EspD is critical for the virulence-mediating ESX-1 secretion system inMycobacterium tuberculosis. J. Bacteriol. 194 884 - 893    DOI : 10.1128/JB.06417-11
Cho SH , Goodlett D , Franzblau S 2006 ICAT-based comparative proteomic analysis of non-replicating persistentMycobacterium tuberculosis. Tuberculosis (Edinb.) 86 445 - 460    DOI : 10.1016/
Cho YS , Dobos KM , Prenni J , Yang H , Hess A , Andersen P 2013 Deciphering the proteome of thein vivodiagnostic reagent “purified protein derivative” fromMycobacterium tuberculosis. Proteomics 12 979 - 991    DOI : 10.1002/pmic.201100544
Chopra P , Singh A , Koul A , Ramachandran S , Drlica K , Tyagi AK , Singh Y 2003 Cytotoxic activity of nucleoside diphosphate kinase secreted fromMycobacterium tuberculosis. Eur. J. Biochem. 270 625 - 634    DOI : 10.1046/j.1432-1033.2003.03402.x
Comstock GW 1975 Frost revisited: the modern epidemiology of tuberculosis. Am. J. Epidemiol. 101 363 - 382
Daffé M , Etienne G 1999 The capsule ofMycobacterium tuberculosisand its implications for pathogenicity. 79 153 - 169
Daniel TM , Janicki BW 1978 Mycobacterial antigens: a review of their isolation, chemistry, and immunological properties. Microbiol. Rev. 42 84 - 113
Darwin KH , Lin G , Chen Z , Li H , Nathan CF 2005 Characterization of aMycobacterium tuberculosisproteasomal ATPase homologue. Mol. Microbiol. 55 561 - 571    DOI : 10.1111/j.1365-2958.2004.04403.x
De Souza GA , Fortuin S , Aguilar D , Pando RH , McEvoy CRE , van Helden PD 2010 Using a label-free proteomics method to identify differentially abundant proteins in closely related hypo- and hypervirulent clinicalMycobacterium tuberculosisBeijing isolates. Mol. Cell. Proteomics 9 2414 - 2423    DOI : 10.1074/mcp.M900422-MCP200
De Souza GA , Leversen NA , Målen H , Wiker HG 2011 Bacterial proteins with cleaved or uncleaved signal peptides of the general secretory pathway. J. Proteomics 75 502 - 510    DOI : 10.1016/j.jprot.2011.08.016
Deenadayalan A , Sundaramurthi JC , Raja A 2010 Immunological and proteomic analysis of preparative isoelectric focusing separated culture filtrate antigens ofMycobacterium tuberculosis. Exp. Mol. Pathol. 88 156 - 162    DOI : 10.1016/j.yexmp.2009.11.008
Drumm JE , Mi K , Bilder P , Sun M , Lim J , Bielefeldt-Ohmann H 2009 Mycobacterium tuberculosisuniversal stress protein Rv2623 regulates bacillary growth by ATP-Binding: requirement for establishing chronic persistent infection. PLoS Pathog. 5 e1000460 -    DOI : 10.1371/journal.ppat.1000460
Muñoz-Elías EJ , McKinney JD 2006 Mycobacterium tuberculosisisocitrate lyases 1 and 2 are jointly required forin vivogrowth and virulence. Nat. Med. 11 638 - 644    DOI : 10.1038/nm1252
Ernst JD 2012 The immunological life cycle of tuberculosis. Nat. Rev. Immunol. 12 581 - 591    DOI : 10.1038/nri3259
Ernst JD , Trevejo-Nuñez G , Banaiee N 2007 Genomics and the evolution, pathogenesis, and diagnosis of tuberculosis. J. Clin. Invest. 117 1738 - 1745    DOI : 10.1172/JCI31810
Esparza M , Palomares B , García T , Espinosa P , Zenteno E , Mancilla R 2014 PstS-1, the 38-kDaMycobacterium tuberculosisglycoprotein, is an adhesin, which binds the macrophage mannose receptor and promotes phagocytosis. Scand. J. Immunol. Epub ahead
Espitia C , Laclette JP , Mondrago M , Amador A , Campuzano J , Martens A 1999 The PE-PGRS glycine-rich proteins ofMycobacterium tuberculosis: a new family of fibronectin-binding proteins? Microbiology 145 3487 - 3495    DOI : 10.1099/00221287-145-12-3487
Espitia C , Rodríguez E , Ramón-Luing L , Echeverría-Valencia G , Vallecillo AJ 2012 Host – pathogen interactions in tuberculosis,InCardona PJ (ed.).Understanding Tuberculosis. Analyzing the Origen of Mycobacterium Tuberculosis Pathogenicity. 1st Ed. InTech Rijeka, Croatia 43 - 76
Fay A , Glickman MS 2014 An essential nonredundant role for mycobacterial DnaK in native protein folding. PLoS Genet. 10 e1004516 -    DOI : 10.1371/journal.pgen.1004516
Florczyk MA , Mccue LA , Stack RF , Hauer CR , Mcdonough KA 2001 Identification and characterization of mycobacterial proteins differentially expressed under standing and shaking culture conditions, including Rv2623 from a novel class of putative ATP-binding proteins. Infect. Immun. 69 5777 - 5785    DOI : 10.1128/IAI.69.9.5777-5785.2001
Flores-Villalva S , Suárez-Güemes F , Espitia C , Whelan AO , Vordermeier M , Gutiérrez-Pabello JA 2012 Specificity of the tuberculin skin test is modified by use of a protein cocktail containing ESAT-6 and CFP-10 in cattle naturally infected withMycobacterium bovis. Clin. Vaccine Immunol. 19 797 - 803    DOI : 10.1128/CVI.05668-11
Flynn JL , Chan J 2003 Immune evasion byMycobacterium tuberculosis: living with the enemy. Curr. Opin. Immunol. 15 450 - 455    DOI : 10.1016/S0952-7915(03)00075-X
Forrellad MA , Klepp LI , Gioffré A , Sabio y , García J , Morbidoni HR 2013 Virulence factors of theMycobacterium tuberculosiscomplex. Virulence 4 3 - 66    DOI : 10.4161/viru.22329
Forrellad MA , Bianco MV , Blanco FC , Nuñez J , Klepp LI , Vazquez CL 2013 Study of thein vivorole of Mce2R, the transcriptional regulator of mce2 operon inMycobacterium tuberculosis. BMC Microbiol. 13 200 -    DOI : 10.1186/1471-2180-13-200
Fukui Y , Hirai T , Uchida T , Yoneda M 1965 Extracellular proteins of tubercle bacilli. IV. Alpha and beta antigens as major extracellular protein products and as cellular components of a strain (H37Rv) ofMycobacterium tuberculosis. Biken J. 8 189 - 199
Ganguly N , Siddiqui I , Sharma P 2008 Role ofM. tuberculosisRD-1 region encoded secretory proteins in protective response and virulence. Tuberculosis 88 510 - 517    DOI : 10.1016/
Gehring AJ , Dobos KM , Belisle JT , Harding CV , Boom WH 2004 Mycobacterium tuberculosisLprG (Rv1411c): a novel TLR-2 ligand that inhibits human macrophage class II MHC antigen processing. J. Immunol. 173 2660 - 2668    DOI : 10.4049/jimmunol.173.4.2660
Gehring AJ , Rojas RE , Canaday DH , David L , Harding CV , Boom WH , Lakey DL 2003 TheMycobacterium tuberculosis19-kilodalton lipoprotein inhibits gamma interferon-regulated HLA-DR and FcγR1 on human macrophages through Toll-like receptor 2. Infect Immun. 71 4487 - 4497    DOI : 10.1128/IAI.71.8.4487-4497.2003
Gevaert K , Vandekerckhove J 2000 Protein identification methods in proteomics proteomics. Electrophoresis 21 1145 - 1154
González-Zamorano M , Mendoza-Hernández G , Xolalpa W , Parada C , Vallecillo AJ , Bigi F , Espitia C 2009 Mycobacterium tuberculosisglycoproteomics based on ConAlectin affinity capture of mannosylated proteins. J. Proteome Res. 8 721 - 733    DOI : 10.1021/pr800756a
Goulding CW , Parseghian A , Sawaya MR , Cascio D , Apostol MI , Gennaro ML , Eisenberg D 2002 Crystal structure of a major secreted protein ofMycobacterium tuberculosis-MPT63 at 1.5-A resolution. Protein Sci. 11 2887 - 2893    DOI : 10.1110/ps.0219002
Gu S , Chen J , Dobos KM , Bradbury EM , Belisle JT , Chen X 2003 Comprehensive proteomic profiling of the membrane constituents of aMycobacterium tuberculosisstrain. Mol. Cell. Proteomics 2 1284 - 1296    DOI : 10.1074/mcp.M300060-MCP200
Gunawardena HP , Feltcher ME , Wrobel JA , Gu S , Miriam B , Chen X 2013 Comparison of the membrane proteome of virulentMycobacterium tuberculosisand the attenuatedMycobacterium bovisBCG vaccine strain by label-free quantitative proteomics. J. Proteome Res. 12 5463 - 5474    DOI : 10.1021/pr400334k
Jagirdar J , Zagzag D 1996 Pathology and insights into pathogenesis of tuberculosis,InRom WN, Garay SM (eds.).Tuberculosis. 1st Ed. Little, Brown and Company New York, N.Y. 467 - 482
Jagusztyn-krynicka ELBK , Roszczenko P , Grabowska A 2009 Impact of proteomics on anti-Mycobacterium tuberculosis(MTB) vaccine development. Polish J. Microbiol. 58 281 - 287
Jiang XIN , Zhang W , Gao F , Huang Y , Lv C , Wang H 2006 Comparison of the proteome of isoniazid-resistant and -susceptible strains ofMycobacterium tuberculosis. Microb. Drug Resist. 12 231 - 238    DOI : 10.1089/mdr.2006.12.231
Johnson S , Brusasca P , Spencer JS , Wiker HG , Shashkina E , Kreiswirth B 2001 Characterization of the secreted MPT53 antigen ofMycobacterium tuberculosis. Infect. Immun. 69 5936 - 5939    DOI : 10.1128/IAI.69.9.5936-5939.2001
Jungblut PR , Müller E , Mattow J , Kaufmann SHE 2001 Proteomics reveals open reading frames inMycobacterium tuberculosisH37Rv not predicted by genomics. Infect. Immun. 69 5905 - 5907    DOI : 10.1128/IAI.69.9.5905-5907.2001
Jungblut PR , Schaible UE , Mollenkopf HJ , Zimny-Arndt U , Raupach B , Mattow J 1999 Comparative proteome analysis ofMycobacterium tuberculosisandMycobacterium bovisBCG strains: towards functional genomics of microbial pathogens. Mol. Microbiol. 33 1103 - 1117    DOI : 10.1046/j.1365-2958.1999.01549.x
Kremer L , Maughan WN , Wilson RA , Dover LG , Besra GS 2002 TheM. tuberculosisantigen 85 complex and mycolyltransferase activity. Lett. Appl. Microbiol. 34 233 - 237    DOI : 10.1046/j.1472-765x.2002.01091.x
Kruh NA , Troudt J , Izzo A , Prenni J , Dobos KM 2010 Portrait of a pathogen: theMycobacterium tuberculosisproteomein vivo. PLoS One 5 e13938 -    DOI : 10.1371/journal.pone.0013938
Kulchavenya E 2014 Extrapulmonary tuberculosis: are statistical reports accurate? Ther. Adv. Infect. Dis. 2 61 - 70
Kurtz S , McKinnon KP , Runge MS , Ting JP-Y , Braunstein M 2006 The SecA2 secretion factor ofMycobacterium tuberculosispromotes growth in macrophages and inhibits the host immune response. Infect. Immun. 74 6855 - 6864    DOI : 10.1128/IAI.01022-06
Lee B , Horwitz MA 1995 Identification of Macrophage and Stress-induced Proteins ofMycobacterium tuberculosis. J. Clin. Invest. 96 245 - 249    DOI : 10.1172/JCI118028
Lew JM , Kapopoulou A , Jones LM , Cole ST 2011 TubercuList--10 years after. Tuberculosis 91 1 - 7    DOI : 10.1016/
Målen H , Berven FS , Fladmark KE , Wiker HG 2007 Comprehensive analysis of exported proteins fromMycobacterium tuberculosisH37Rv. Proteomics 7 1702 - 1718    DOI : 10.1002/pmic.200600853
Målen H , Berven FS , Søfteland T , Arntzen MØ , D’Santos CS , De Souza GA , Wiker HG 2008 Membrane and membrane-associated proteins in Triton X-114 extracts ofMycobacterium bovisBCG identified using a combination of gel-based and gel-free fractionation strategies. Proteomics 8 1859 - 1870    DOI : 10.1002/pmic.200700528
Målen H , Pathak S , Søfteland T , de Souza GA , Wiker HG 2010 Definition of novel cell envelope associated proteins in Triton X-114 extracts ofMycobacterium tuberculosisH37Rv. BMC Microbiol. 10 132 -    DOI : 10.1186/1471-2180-10-132
Målen H , De Souza G a , Pathak S , Søfteland T , Wiker HG 2011 Comparison of membrane proteins ofMycobacterium tuberculosisH37Rv and H37Ra strains. BMC Microbiol. 11 18 -    DOI : 10.1186/1471-2180-11-18
Manca C , Lyashchenko K , Colangeli R , Gennaro ML , Manca C , Lyashchenko K 1997 MTC28, a novel 28-kilodalton proline-rich secreted antigen specific for theMycobacterium tuberculosiscomplex. Infect. Immun. 65 4951 - 4957
Mattow J , Jungblut PR , Schaible UE , Mollenkopf H , Lamer S , Zimny-arndt U 2001 Identification of proteins fromMycobacterium tuberculosismissing in attenuatedMycobacterium bovisBCG strains. Electrophoresis 22 2936 - 2946
Mattow J , Schaible UE , Schmidt F , Hagens K , Siejak F , Brestrich G 2003 Comparative proteome analysis of culture supernatant proteins from virulentMycobacterium tuberculosisH37Rv and attenuatedM. bovisBCG Copenhagen. Electrophoresis 24 3405 - 3420    DOI : 10.1002/elps.200305601
Mattow J , Siejak F , Hagens K , Becher D , Albrecht D , Krah A 2006 Proteins unique to intraphagosomally grownMycobacterium tuberculosis. Proteomics 6 2485 - 2494    DOI : 10.1002/pmic.200500547
Mawuenyega KG , Forst CV , Dobos KM , Belisle JT , Chen J , Bradbury EM 2005 Mycobacterium tuberculosisfunctional network analysis by global subcellular protein profiling. Mol. Biol. Cell 16 396 - 404    DOI : 10.1091/mbc.E04-04-0329
Monahan IM , Betts J , Banerjee DK , Butcher PD 2001 Differential expression of mycobacterial proteins following phagocytosis by macrophages. Microbiology 147 459 - 471    DOI : 10.1099/00221287-147-2-459
Nagai S , Wiker HG , Harboe M , Kinomoto M 1991 Isolation and partial characterization of major protein antigens in the culture fluid ofMycobacterium tuberculosis. Infect. Immun. 59 372 - 382
O’Garra A , Redford PS , McNab FW , Bloom CI , Wilkinson RJ , Berry MPR 2013 The immune response in tuberculosis. Annu. Rev. Immunol. 31 475 - 527    DOI : 10.1146/annurev-immunol-032712-095939
Pai RK , Convery M , Hamilton TA , Boom WH , Harding CV 2003 Inhibition of IFN- -induced class II transactivator expression by a 19-kDa lipoprotein fromMycobacterium tuberculosis: a potential mechanism for immune evasion. J. Immunol. 171 175 - 184    DOI : 10.4049/jimmunol.171.1.175
Parish T 2003 The senX3-regX3 two-component regulatory system ofMycobacterium tuberculosisis required for virulence. Microbiology 149 1423 - 1435    DOI : 10.1099/mic.0.26245-0
Patel VJ , Thalassinos K , Slade SE , Connolly JB , Crombie A , Murrell JC , Scrivens JH 2009 A comparison of labeling and label-free mass spectrometry-based proteomics approaches. J. Proteome Res. 8 3752 - 3759    DOI : 10.1021/pr900080y
Pecora ND , Gehring AJ , Canaday DH , Boom WH , Harding CV 2006 Mycobacterium tuberculosisLprA is a lipoprotein agonist of TLR2 that regulates innate immunity and APC function. J. Immunol. 177 422 - 429    DOI : 10.4049/jimmunol.177.1.422
Pethe K , Alonso S , Biet F , Delogu G , Brennan MJ , Locht C , Menozzi FD 2001 The heparin-binding haemagglutinin ofM. tuberculosisis required for extrapulmonary dissemination. Nature 412 190 - 194    DOI : 10.1038/35084083
Pieters J 2008 Mycobacterium tuberculosisand the macrophage: maintaining a balance. Cell Host Microbe. 3 399 - 407    DOI : 10.1016/j.chom.2008.05.006
Prasad TSK , Verma R , Kumar S , Nirujogi RS , Sathe GJ , Madugundu AK 2013 Proteomic analysis of purified protein derivative ofMycobacterium tuberculosis. Clin. Proteomics 10 8 -    DOI : 10.1186/1559-0275-10-8
Puffer RR , Steward HC , Gass RS 1945 Tuberculosis in house- hold contacts associates: the inuence of age and relationship. Am. Rev. Tuberc 52 89 - 103
Papavinasasundaram KG , Speight RA , Springer B , Sander P , Böttger EC 2002 The functions of OmpATb, a pore-forming protein ofMycobacterium tuberculosis. Mol. Microbiol. 46 191 - 201    DOI : 10.1046/j.1365-2958.2002.03152.x
Reddy MCM , Kuppan G , Shetty ND , Owen JL , Ioerger TR , Sacchettini JC 2008 Crystal structures ofMycobacterium tuberculosisS-adenosyl-L-homocysteine hydrolase in ternary complex with substrate and inhibitors. Protein Sci. 17 2134 - 2144    DOI : 10.1110/ps.038125.108
Rifat D , Belchis DA , Karakousis PC 2014 senX3- independent contribution of regX3 toMycobacterium tuberculosisvirulence. BMC Microbiol. 14 265 -    DOI : 10.1186/s12866-014-0265-8
Roberts MM , Coker AR , Fossati G , Coates ARM , Wood SP , Mascagni P 2003 Mycobacterium tuberculosischaperonin 10 heptamers self-associate through their biologically active loops. J. Bacteriol. 185 4172 - 4185    DOI : 10.1128/JB.185.14.4172-4185.2003
Rohde K , Yates RM , Purdy GE , Russell DG 2007 Mycobacterium tuberculosisand the environment within the phagosome. 219 37 - 54
Rosenkrands I , King A , Weldingh K , Moniatte M , Moertz E , Andersen P 2000 Towards the proteome ofMycobacterium tuberculosis. Electrophoresis 21 3740 - 3756
Rosenkrands I , Slayden RA , Crawford J , Aagaard C , Barry CE III , Andersen P 2002 Hypoxic response ofMycobacterium tuberculosisstudied by metabolic labeling and proteome analysis of cellular and extracellular proteins. J. Bacteriol. 184 3485 - 3491    DOI : 10.1128/JB.184.13.3485-3491.2002
Rosenkrands I , Weildingh K , Jacobsen S , Hansen VC , Florio W , Gianetri I , Andersen P 2000 Mapping and identification ofMycobacterium tuberculosisproteins by two-dimensional gel electrophoresis, microsequencing and immunodetection. Electrophoresis 21 935 - 948
Russell DG 2007 Who puts the tubercle in tuberculosis? Nat. Rev. Microbiol. 5 39 - 47    DOI : 10.1038/nrmicro1538
Russell RB , Eggleston DS 2000 New roles for structure in biology and drug discovery. Nat. Struct. Biol. 7 928 - 930    DOI : 10.1038/80691
Ryndak M , Wang S , Smith I 2008 PhoP, a key player inMycobacterium tuberculosisvirulence. Trends Microbiol. 16 528 - 534    DOI : 10.1016/j.tim.2008.08.006
Sachdeva P , Misra R , Tyagi AK , Singh Y 2010 The sigma factors ofMycobacterium tuberculosis: regulation of the regulators. FEBS J. 277 605 - 626    DOI : 10.1111/j.1742-4658.2009.07479.x
Sajid A , Arora G , Gupta M , Singhal A , Chakraborty K , Nandicoori VK , Singh Y 2011 Interaction ofMycobacterium tuberculosiselongation factor Tu with GTP is regulated by phosphorylation. J. Bacteriol. 193 5347 - 5358    DOI : 10.1128/JB.05469-11
Shukla S , Richardson ET , Athman JJ , Shi L , Wearsch PA , McDonald D 2014 Mycobacterium tuberculosislipoprotein LprG binds lipoarabinomannan and determines its cell envelope localization to control phagolysosomal fusion. PLoS Pathog. 10 e1004471 -    DOI : 10.1371/journal.ppat.1004471
Sidders B , Pirson C , Hogarth PJ , Hewinson RG , Stoker NG , Vordermeier HM , Ewer K 2008 Screening of highly expressed mycobacterial genes identifies Rv3615c as a useful differential diagnostic antigen for theMycobacterium tuberculosiscomplex. Infect. Immun. 76 3932 - 3939    DOI : 10.1128/IAI.00150-08
Siddiqui KF , Amir M , Agrewala JN 2011 Understanding the biology of 16 kDa antigen ofMycobacterium tuberculosis: scope in diagnosis, vaccine design and therapy. Crit. Rev. Microbiol. 37 349 - 357    DOI : 10.3109/1040841X.2011.606425
Singhal N , Sharma P , Kumar M , Joshi B , Bisht D 2012 Analysis of intracellular expressed proteins ofMycobacterium tuberculosisclinical isolates. Proteome Sci. 10 14 -    DOI : 10.1186/1477-5956-10-14
Sinha S , Kosalai K , Arora S , Namane A , Sharma P , Gaikwad AN 2005 Immunogenic membraneassociated proteins ofMycobacterium tuberculosisrevealed by proteomics Microbiology 151 2411 - 2419    DOI : 10.1099/mic.0.27799-0
Smith CV , Huang C , Miczak A , Russell DG , Sacchettini JC , Höner zu Bentrup K 2003 Biochemical and structural studies of malate synthase fromMycobacterium tuberculosis. J. Biol. Chem. 278 1735 - 1743    DOI : 10.1074/jbc.M209248200
Sonnenberg MG , Belisle JT 1997 Definition ofMycobacterium tuberculosisculture filtrate proteins by two-dimensional polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and electrospray mass spectrometry. Infect. Immun. 65 4515 - 4524
Sørensen AL , Nagai S , Houen G , Andersen P , Andersen AB 1995 Purification and characterization of a low-molecularmass T-cell antigen secreted byMycobacterium tuberculosis. Infect. Immun. 63 1710 - 1717
Sreejit G , Ahmed A , Parveen N , Jha V , Valluri VL , Ghosh S , Mukhopadhyay S 2014 The ESAT-6 protein ofMycobacterium tuberculosisinteracts with beta-2-microglobulin (β2M) affecting antigen presentation function of macrophage. PLoS Pathog. 10 e1004446 -    DOI : 10.1371/journal.ppat.1004446
Starck J , Källenius G , Marklund B-I , Andersson DI , Akerlund T 2004a Comparative proteome analysis ofMycobacterium tuberculosisgrown under aerobic and anaerobic conditions. Microbiology 150 3821 - 3829    DOI : 10.1099/mic.0.27284-0
Vilchèze C , Molle V , Carrère-Kremer S , Leiba J , Mourey L , Shenai S 2014 Phosphorylation of KasB regulates virulence and acid-fastness inMycobacterium tuberculosis. PLoS Pathog. 10 e1004115 -    DOI : 10.1371/journal.ppat.1004115
Vordermeier M , Jones GJ , Whelan AO 2011 DIVA reagents for bovine tuberculosis vaccines in cattle. Expert Rev. Vaccines 10 1083 - 1091    DOI : 10.1586/erv.11.22
Wang Z , Potter BM , Gray AM , Sacksteder KA , Geisbrecht BV , Laity JH 2007 The solution structure of antigen MPT64 from Mycobacterium tuberculosisdefines a new family of beta-grasp proteins. J. Mol. Biol. 366 375 - 381    DOI : 10.1016/j.jmb.2006.11.039
Weldingh K , Rosenkrands I , Jacobsen S , Birk P , Elhay MJ , Andersen P 1998 Two-dimensional electrophoresis for analysis ofMycobacterium tuberculosisculture filtrate and purification and characterization of six novel proteins. Infect. Immun. 66 3492 - 3500
Wiker HG 2009 MPB70 and MPB83--major antigens ofMycobacterium bovis. Scand. J. Immunol. 69 492 - 499    DOI : 10.1111/j.1365-3083.2009.02256.x
Wilson RA , Maughan WN , Kremer L , Besra GS , Fütterer K 2004 The structure ofMycobacterium tuberculosisMPT51 (FbpC1) defines a new family of non-catalytic alpha/beta hydrolases. J. Mol. Biol. 335 519 - 530    DOI : 10.1016/j.jmb.2003.11.001
World Health Organization 2013 Global Tuberculosis Report. WHO France
Wu L , Zhang M , Sun M , Jia B , Wang X 2011 Humoural immune responses to a recombinant 16-kDa-38-kDa--ESAT-6Mycobacterial Antigenin tuberculosis. J. Int. Med. Res. 39 514 - 521    DOI : 10.1177/147323001103900219
Xiong Y , Chalmers MJ , Gao FP , Cross TA , Marshall AG 2005 Identification ofMycobacterium tuberculosisH37Rv integral membrane proteins by one-dimensional gel electrophoresis and liquid chromatography electrospray ionization tandem mass spectrometry. J. Proteome Res. 4 855 - 861    DOI : 10.1021/pr0500049
Xolalpa W , Vallecillo AJ , Lara M , Mendoza-Hernandez G , Comini M , Spallek R , Singh M , Espitia C 2007 Identification of novel bacterial plasminogen-binding proteins in the human pathogenMycobacterium tuberculosis. Proteomics 7 3332 - 3341    DOI : 10.1002/pmic.200600876
Yang H , Kruh-Garcia NA , Dobos KM 2012 Purified protein derivatives of tuberculin - past, present, and future. FEMS Immunol. Med Microbiol. 66 273 - 280    DOI : 10.1111/j.1574-695X.2012.01002.x
Yang H , Troudt J , Grover A , Arnett K , Lucas M , Cho YS 2011 Three protein cocktails mediate delayed-type hypersensitivity responses indistinguishable from that elicited by purified protein derivative in the guinea pig model ofMycobacterium tuberculosisinfection. Infect. Immun. 79 716 - 723    DOI : 10.1128/IAI.00486-10
Young DB , Kaufmann SHE , Hermans PWM 1992 Mycobacterial protein antigens: a compilation. Mol. Microbiol. 6 133 - 145    DOI : 10.1111/j.1365-2958.1992.tb01994.x
Zheng J , Liu L , Wei C , Leng W , Yang J , Li W 2012 A comprehensive proteomic analysis ofMycobacterium bovisbacillus Calmette-Guérin using high resolution Fourier transform mass spectrometry. J. Proteomics 77 357 - 371    DOI : 10.1016/j.jprot.2012.09.010
Zheng J , Ren X , Wei C , Yang J , Hu Y , Liu L , Xu X , Wang J , Jin Q 2013 Analysis of the secretome and identification of novel constituents from culture filtrate of bacillus Calmette-Guerin using high-resolution mass spectrometry. Mol. Cell. Proteomics 12 2081 - 2095    DOI : 10.1074/mcp.M113.027318