Identification, expression and serological evaluation of the recombinant ATP synthase beta subunit of Mycoplasma pneumoniae

Nuyttens, Hélène; Cyncynatus, Camille; Renaudin, H.; Pereyre, Sabine; Bebear, Cécile

doi:10.1186/1471-2180-10-216

Cited by 12 publications

(7 citation statements)

References 43 publications

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“…Using proteome analysis, PDHB was identified as an antigen that reacts with M. pneumoniae-positive patient sera (Nuyttens et al, 2010). The interaction of PDHB with plasminogen and the previously described binding to human fibronectin (Dallo et al, 2002) show that this molecule belongs to the group of proteins mediating contact to more than one ECM component, such as enolase of P. brasiliensis (Donofrio et al, 2009;Nogueira et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

2013

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The obligate pathogenic mycoplasma species Mycoplasma pneumoniae uses a limited but effective repertoire of virulence factors to infect and colonize the human respiratory tract. Besides the development of a unique adhesion complex and the expression of tissue-damaging factors, surface-located glycolytic enzymes and their capacity to bind to components of the human extracellular matrix (ECM) support pathogen-host interactions. Here, we demonstrated that the glycolytic enzymes enolase (Mpn606) and pyruvate dehydrogenase subunit B (Mpn392; PDHB) of M. pneumoniae show concentration-dependent binding to human plasminogen. Monospecific polyclonal antisera against both recombinant proteins reduced the binding to plasminogen significantly. The surface location of PDHB but not of enolase was demonstrated using Triton X fractionation of M. pneumoniae total protein content, membrane fractionation, colony blotting, mild proteolysis of mycoplasma cells, and immunofluorescence tests. To characterize the binding site of plasminogen in surface-displaced PDHB, the mycoplasmal protein was separated into four recombinant proteins followed by investigation of the binding behaviour of peptides that overlap the protein part interacting with plasminogen. Spot analysis resulted in a novel region of 12 amino acids (FPAMFQIFTHAA, position 91 to 102 of PDHB), which is responsible exclusively for binding of human plasminogen and also interacts in a dose-dependent manner with this host protein. The data indicate that the plasminogen-binding enzymes enolase and especially the surface-associated PDHB may contribute to the pathogenesis of M. pneumoniae infections.

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Section: Discussionmentioning

confidence: 99%

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

2013

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“…An initial pilot study indicated that vaccination of BALB/c mice with LAMPs resulted in VED after challenge with virulent Mp. We performed proteomic analysis to reveal lipoproteins, elongation factors, chaperones and chaperonins, and cytadherence proteins (previously found to induce strong antibody responses [15][16][17] ) as the most abundant immunogenic and antigenic components of the LAMP fraction ( Fig. 1a-f).…”

Section: Resultsmentioning

confidence: 99%

Lipid moieties of Mycoplasma pneumoniae lipoproteins are the causative factor of vaccine-enhanced disease

et al. 2020

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Vaccine-enhanced disease (VED) occurs as a result of vaccination followed by infection with virulent Mycoplasma pneumoniae. To date VED has prevented development of an efficacious vaccine against this significant human respiratory pathogen. Herein we report that vaccination of BALB/c mice with M. pneumoniae lipid-associated membrane proteins (LAMPs) induces lung lesions consistent with exacerbated disease following challenge, without reducing bacterial loads. Removal of lipid moieties from LAMPs prior to vaccination eliminates VED and reduces bacterial loads after infection. Collectively, these data indicate that lipid moieties of lipoproteins are the causative factors of M. pneumoniae VED.npj Vaccines (2020) 5:31 ; https://doi.

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“…Nuyttens et al . () identified the M. pneumoniae ATP synthase beta‐subunit (AtpD) by serologic proteome analysis, and their results suggest that AtpD can be used as an antigen for the immunodiagnosis of early and acute M. pneumoniae infection in association with adhesin P1. The above research indicated P1 proteins as a promising specific antigen candidate for the diagnosis of M. pneumoniae infection.…”

Section: Discussionmentioning

confidence: 99%

“…Nuyttens et al . () reported the sensitivity and specificity of the C‐terminal part of P1 protein were 69 and 90%, respectively, when tested for IgG antibodies in 54 patients and 86 controls (Nuyttens et al ., ). In our study, two epitopes were used for serodiagnosis, with rP1‐513 having a higher sensitivity than rP1‐534 for IgG and IgM antibodies.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of P1 adhesin epitopes for the serodiagnosis ofMycoplasma pneumoniaeinfections

Xue

Cao

Wang

et al. 2013

FEMS Microbiol Lett

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Most glycolipid antigens used for serological tests of Mycoplasma pneumoniae are not M. pneumonia-specific, and can cross-react with other microorganism antigens and body tissues, resulting in false positives. It is important to identify M. pneumonia-specific antigen(s) for serological testing and correct diagnosis. Two epitopes, rP1-534 and rP1-513, of P1 adhesin predicted by bioinformatics were successfully expressed and purified, and could be recognized by serum samples from M. pneumoniae-infected patients and His tag antibodies by Western blot. There was no cross-reactivity between the anti-recombinant proteins serum and other respiratory antigens. A total of 400 patients were investigated, their respiratory specimens tested by PCR, and sera tested by a commercial test kit; 56 with positive sera and positive respiratory specimens were designated as standard positive serum and 63 patients were designated as standard negative serum. The purified recombinant proteins were used as a combination of antigens or separately to test the serum. Serological test demonstrated that rP1-513 of the C terminal of P1 adhesin is a new candidate antigen with greater sensitivity and specificity for IgG and IgM serodiagnosis of M. pneumoniae-infected patients. The results confirmed that rP1-513 could be a useful new antigen for the immunodiagnosis of M. pneumoniae infection.

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Identification, expression and serological evaluation of the recombinant ATP synthase beta subunit of Mycoplasma pneumoniae

Cited by 12 publications

References 43 publications

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

Lipid moieties of Mycoplasma pneumoniae lipoproteins are the causative factor of vaccine-enhanced disease

Evaluation of P1 adhesin epitopes for the serodiagnosis ofMycoplasma pneumoniaeinfections

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