Biochemistry of the Cell Envelope of<i>Mycobacterium tuberculosis</i>

Crick, Dean C.; Quadri, Luis; Brennan, Patrick J.

doi:10.1002/9783527611614.ch1

Cited by 7 publications

(3 citation statements)

References 114 publications

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“…Proteins localized to the membrane of M. tuberculosis play critical roles in vital cell processes including nutrient transport, cell-wall synthesis, energy metabolism, and signal transduction. − Additionally, mycobacterial membrane proteins can elicit immune responses, making the membrane proteomes of M. tuberculosis and BCG of significant interest for vaccination and diagnostic studies . Initial efforts to identify the M. tuberculosis and BCG membrane proteome used 2D-GE; however, the high insolubility of membrane proteins poses a significant technical challenge for 2D-GE and limits the numbers of proteins that can be identified. , Significantly better protein identification coverage was subsequently obtained when membrane proteins were solubilized and pre-separated by 1D SDS-PAGE, followed by LC–MS/MS analysis of trypsin digested gel slices comprising the entire sample .…”

Section: Introductionmentioning

confidence: 99%

Comparison of the Membrane Proteome of Virulent Mycobacterium tuberculosis and the Attenuated Mycobacterium bovis BCG Vaccine Strain by Label-Free Quantitative Proteomics

et al. 2013

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The Mycobacterium tuberculosis (MTB) 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 (LFQ) proteomics, utilizing the area-under-curve (AUC) of the extracted ion chromatograms (XIC) of peptides obtained from m/z and retention time alignment of MS1 features. With this approach, we obtained relative abundance ratios for 2,203 identified membrane-associated proteins in high confidence. Of these proteins, 294 showed statistically significant differences of at least 2 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.

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Section: Introductionmentioning

confidence: 99%

Comparison of the Membrane Proteome of Virulent Mycobacterium tuberculosis and the Attenuated Mycobacterium bovis BCG Vaccine Strain by Label-Free Quantitative Proteomics

et al. 2013

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“…Membrane proteins of M. tuberculosis play major roles in cell processes including nutrient transport, cell wall synthesis, energy metabolism, and signal transduction . Identification of targets among membrane proteins is critical but challenging in anti‐TB drug development.…”

Section: Discussionmentioning

confidence: 99%

Viability, morphology, and proteome of Mycobacterium smegmatis MSMEG_0319 knockout strain

Sha

Shi

et al. 2016

Proteomics

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Mycobacterium tuberculosis Rv0228, a membrane protein, is predicted as a drug target through computational methods. MSMEG_0319 (MS0319) in Mycobacterium smegmatis mc(2) 155 is the ortholog of Rv0228. To study the effect of MS0319 protein on M. smegmatis, an MS0319 gene knockout strain (ΔMS0319) was generated via a homologous recombination technique in this study. The results showed that the lack of MS0319 protein in mc(2) 155 cells led to the loss of viability at nonpermissive temperature. Scanning electron microscopy and transmission electron microscopy observations showed drastic changes in cellular shape especially cell wall disruption in ΔMS0319 cells. Proteomic analysis of ΔMS0319 cells through LC-MS/MS revealed that 462 proteins had changes in their expressions by lacking MS0319 protein. The M. tuberculosis orthologs of these 462 proteins were found through BLASTp search and functional clustering and metabolic pathway enrichment were performed on the orthologs. The results revealed that most of them were enzymes involved in metabolism of carbohydrates and amino acids, indicating that Rv0228 played an important role in cellular metabolism. All these results suggested Rv0228 as a potential target for development of antituberculosis drugs.

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“…Many pathogenic Mycobacterium such as Mycobacterium tuberculosis, Mycobacterium leprae, Mycobacterium avium, Mycobacterium ulcerans, Mycobacterium bovis, and Mycobacterium marinum are generally slow-growing; however, a strict relationship between pathogenicity and growth remains to be established . In addition to these slow growth patterns, mycobacteria have developed an unusual cell envelope, which is thicker than in most other bacteria, and distinctively rich in mycolic acids and their derivatives. − Consistent with these characteristics, almost 10% of the genes encoded by Mycobacterium produce proteins involved in the metabolism of lipids for either cell wall synthesis or β-oxidation . This underscores the importance of understanding lipid metabolism in mycobacteria, with the potential for development of new drugs targeting these essential enzymes.…”

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confidence: 99%

Mycobacteria Encode Active and Inactive Classes of TesB Fatty-Acyl CoA Thioesterases Revealed through Structural and Functional Analysis

et al. 2017

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Mycobacteria contain a large number of highly divergent species and exhibit unusual lipid metabolism profiles, believed to play important roles in immune invasion. Thioesterases modulate lipid metabolism through the hydrolysis of activated fatty-acyl CoAs; multiple copies are present in mycobacteria, yet many remain uncharacterized. Here, we undertake a comprehensive structural and functional analysis of a TesB thioesterase from Mycobacterium avium (MaTesB). Structural superposition with other TesB thioesterases reveals that the Asp active site residue, highly conserved across a wide range of TesB thioesterases, is mutated to Ala. Consistent with these structural data, the wild-type enzyme failed to hydrolyze an extensive range of acyl-CoA substrates. Mutation of this residue to an active Asp residue restored activity against a range of medium-chain length fatty-acyl CoA substrates. Interestingly, this Ala mutation is highly conserved across a wide range of Mycobacterium species but not found in any other bacteria or organism. Our structural homology analysis revealed that at least one other TesB acyl-CoA thioesterase also contains an Ala residue at the active site, while two other Mycobacterium TesB thioesterases harbor an Asp residue at the active site. The inactive TesBs display a common quaternary structure that is distinct from that of the active TesB thioesterases. Investigation of the effect of expression of either the catalytically active or inactive MaTesB in Mycobacterium smegmatis exposed, to the best of our knowledge, the first genotype-phenotype association implicating a mycobacterial tesB gene. This is the first report that mycobacteria encode active and inactive forms of thioesterases, the latter of which appear to be unique to mycobacteria.

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Biochemistry of the Cell Envelope ofMycobacterium tuberculosis

Cited by 7 publications

References 114 publications

Comparison of the Membrane Proteome of Virulent Mycobacterium tuberculosis and the Attenuated Mycobacterium bovis BCG Vaccine Strain by Label-Free Quantitative Proteomics

Comparison of the Membrane Proteome of Virulent Mycobacterium tuberculosis and the Attenuated Mycobacterium bovis BCG Vaccine Strain by Label-Free Quantitative Proteomics

Viability, morphology, and proteome of Mycobacterium smegmatis MSMEG_0319 knockout strain

Mycobacteria Encode Active and Inactive Classes of TesB Fatty-Acyl CoA Thioesterases Revealed through Structural and Functional Analysis

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