A Comprehensive Proteome of<i>Mycoplasma genitalium</i>

Párraga-Niño, Noemí; Colomé-Calls, Núria; Canals, Françesc; Querol, Enrique; Ferrer-Navarro, Mario

doi:10.1021/pr300084c

Cited by 24 publications

(21 citation statements)

References 60 publications

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“…Interestingly, our study validates the findings of a previous study which identified pyruvate dehydrogenase E1 α chain and the protein encoded by MG_328 as phosphoproteins of M. genitalium [47]. Recently, both these proteins were found on the surface of M. genitalium [48], thus suggesting the possibility that they can play a role in M. genitalium -host interaction. What is intriguing in this study, however, is the reduced phosphorylation of a protein (PDH) in a phosphatase (MG208) deficient mutant of M. genitalium .…”

Section: Resultssupporting

confidence: 90%

A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence

et al. 2013

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BackgroundBacterial signal transduction systems like two component system (TCS) and Serine/Threonine kinase (STK) and Serine/Threonine phosphatase (STP) play important roles in the virulence and pathogenesis of bacterial pathogens. Mycoplasma genitalium, a mollicute that causes the urogenital diseases urethritis and cervicitis in men and women, respectively, is a pathogen which lacks TCS but possesses STK/STP. In this study, we investigated the biochemical and virulence properties of an STP protein encoded by the gene MG_207 of this species.ResultsWe overexpressed MG207 in Escherichia coli overexpression system as a recombinant His10MG207 protein and purified it with affinity chromatography. This recombinant protein readily hydrolyzed the substrate p-nitrophenyl phosphate (pNPP) in a dose-dependent manner. Additional studies using synthetic peptides as substrates revealed that the recombinant protein was able to hydrolyze the threonine phosphate. Further, a transposon insertion mutant strain of M. genitalium (TIM207) that lacks the protein MG207 showed differentially phosphorylated proteins when compared to the wild type G37 strain. Mass spectrometry revealed that some of the key proteins differentially phosphorylated in TIM207 strain were putative cytoskeletal protein encoded by the gene MG_328 and pyruvate dehydrogenase E1 α chain encoded by the gene MG_274. In addition, TIM207 was noticed to be less cytotoxic to HeLa cells and this correlated with the production of less hydrogen peroxide by this strain. This strain was also less efficient in inducing the differentiation of THP-1 cell line as compared to wild type M. genitalium.ConclusionsThe results of the study suggest that MG207 is an important signaling protein of M. genitalium and its presence may be crucial for the virulence of this species.

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Section: Resultssupporting

confidence: 90%

A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence

et al. 2013

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“…In this study, a total of 404 of the 795 M. suis KI_3806 proteins corresponding to 50.8% of all encoded proteins were identified (Supporting Information Files 3 and 4). This identification ratio is in accordance with GeLC‐MS/MS protein identification levels for other cultivable mycoplasmas . The proteins were distributed as follows: 316 proteins in the M. suis enriched fraction (method A), 229 in the buffer‐soluble fraction (method B), and 268 proteins in the buffer‐insoluble fraction (method C).…”

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confidence: 80%

Updating the proteome of the uncultivable hemotrophicMycoplasma suisin experimentally infected pigs

et al. 2016

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Mycoplasma suis belongs to the hemotrophic mycoplasmas that are associated with acute and chronic anemia in a wide range of livestock and wild animals. The inability to culture M. suis in vitro has hindered its characterization at the molecular level. Since the publication of M. suis genome sequences in 2011 only one proteome study has been published. Aim of the presented study was to significantly extend the proteome coverage of M. suis strain KI_3806 during acute infection by applying three different protein extraction methods followed by 1D SDS-PAGE and LC-MS/MS. A total of 404 of 795 M. suis KI_3806 proteins (50.8%) were identified. Data analysis revealed the expression of 83.7% of the predicted ORFs with assigned functions but also highlights the expression of 179 of 523 (34.2%) hypothetical proteins with unknown functions. Computational analyses identified expressed membrane-associated hypothetical proteins that might be involved in adhesion or host-pathogen interaction. Furthermore, analyses of the expressed proteins indicated the existence of a hexose-6-phosphate-transporter and an ECF transporter. In conclusion, our proteome study provides a further step toward the elucidation of the unique life cycle of M. suis and the establishment of an in vitro culture. All MS data have been deposited in the ProteomeXchange with identifier PXD002294 (http://proteomecentral.proteomexchange.org/dataset/PXD002294).

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“…M. bovis MAPs were fractionated using TX-114 as previously described [48] with minor modifications. In brief, M. bovis HB0801 pellet was re-suspended in PBS containing 4% TX-114 (Sigma) and 1 mM PMSF (Sigma) and kept for 3-5h at 4°C after scraping.…”

Section: Methodsmentioning

confidence: 99%

Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

et al. 2016

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A lack of knowledge regarding the antigenic properties of Mycoplasma bovis proteins prevents the effective control of bovine infections using immunological approaches. In this study, we detected and characterized a specific and sensitive M. bovis diagnostic biomarker. After M. bovis total proteins and membrane fractions were separated with two dimensional gel electrophoresis, proteins reacting with antiserawere detected using MALDI-TOF MS. Thirty-nine proteins were identified, 32 of which were previously unreported. Among them, immunoinformatics predicted eight antigens, encoded by Mbov_0106, 0116, 0126, 0212, 0275, 0579, 0739, and 0789, to have high immunological value. These genes were expressed in E. coli after mutagenesis of UGA to UGG using overlap extension PCR. A lipoprotein, MbovP579, encoded by a functionally unknown gene, was a sensitive and specific antigen for detection of antibodies in sera from both M. bovis-infected and vaccinated cattle. The specificity of MbovP579 was confirmed by its lack of cross-reactivity with other mycoplasmas, including Mycoplasma agalactiae. An iELISA based on rMbovP579 detected seroconversion 7 days post-infection (dpi). The ELISA had sensitivity of 90.2% (95% CI: 83.7%, 94.3%) and a specificity of 97.8% (95% CI: 88.7%, 99.6%) with clinical samples. Additional comparative studies showed that both diagnostic and analytic sensitivities of the ELISA were higher than those of a commercially available kit (p<0.01). We have thus detected and characterized the novel antigen, MbovP579, and established an rMbovP579-based ELISA as a highly sensitive and specific method for the early diagnosis of M. bovis infection.

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A Comprehensive Proteome ofMycoplasma genitalium

Cited by 24 publications

References 60 publications

A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence

A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence

Updating the proteome of the uncultivable hemotrophicMycoplasma suisin experimentally infected pigs

Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

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