Identification of membrane-associated proteins with pathogenic potential expressed by Corynebacterium pseudotuberculosis grown in animal serum

Filho, José Tadeu Raynal Rocha; Bastos, Bruno Lopes; Vilas-Boas, Priscilla Carolinne Bagano; Sousa, Thiago de Jesus; Costa-Silva, Marcos; Sá, Maria da Conceição Aquino de; Portela, Ricardo Wagner; Moura-Costa, Lília Ferreira de; Azevedo, Vasco; Meyer, Roberto

doi:10.1186/s13104-018-3180-5

Cited by 9 publications

(10 citation statements)

References 33 publications

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“…Only five secreted and four cell wall‐localized proteins were detectable by MS analysis indicating that the membrane samples were largely free of extracellular proteins. In contrast, numerous cytoplasmic proteins were found to be associated with the membrane preparations as is expected for these types of studies (Cathro, McCarthy, Hoffmann, & Zilm, ; Fantappiè et al, ; Gunawardena et al, ; Halbedel et al, ; Price et al, ; Raynal et al, ; Zanen et al, ). The largest number of proteins was identified for the ∆yidC1 strain, and smallest number for the ∆ffh/yidC1 mutant.…”

Section: Resultsmentioning

confidence: 58%

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Membrane proteomic analysis reveals overlapping and independent functions of Streptococcus mutans Ffh, YidC1, and YidC2

Mishra

Crowley

Wright

et al. 2019

Molecular Oral Microbiology

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A comparative proteomic analysis was utilized to evaluate similarities and differences in membrane samples derived from the cariogenic bacterium Streptococcus mutans, including the wild‐type strain and four mutants devoid of protein translocation machinery components, specifically ∆ffh, ∆yidC1, ∆yidC2, or ∆ffh/yidC1. The purpose of this work was to determine the extent to which the encoded proteins operate individually or in concert with one another and to identify the potential substrates of the respective pathways. Ffh is the principal protein component of the signal recognition particle (SRP), while yidC1 and yidC2 are dual paralogs encoding members of the YidC/Oxa/Alb family of membrane‐localized chaperone insertases. Our results suggest that the co‐translational SRP pathway works in concert with either YidC1 or YidC2 specifically, or with no preference for paralog, in the insertion of most membrane‐localized substrates. A few instances were identified in which the SRP pathway alone, or one of the YidCs alone, appeared to be most relevant. These data shed light on underlying reasons for differing phenotypic consequences of ffh, yidC1 or yidC2 deletion. Our data further suggest that many membrane proteins present in a ∆yidC2 background may be non‐functional, that ∆yidC1 is better able to adapt physiologically to the loss of this paralog, that shared phenotypic properties of ∆ffh and ∆yidC2 mutants can stem from impacts on different proteins, and that independent binding to ribosomal proteins is not a primary functional activity of YidC2. Lastly, genomic mutations accumulate in a ∆yidC2 background coincident with phenotypic reversion, including an apparent W138R suppressor mutation within yidC1.

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Section: Resultsmentioning

confidence: 58%

“… Transmembrane domains (TMD) were predicted using the TMHMM 2.0 transmembrane helix prediction tool (Raynal et al, ). …”

Section: Resultsmentioning

confidence: 99%

Membrane proteomic analysis reveals overlapping and independent functions of Streptococcus mutans Ffh, YidC1, and YidC2

Mishra

Crowley

Wright

et al. 2019

Molecular Oral Microbiology

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“…Few methods use recombinant proteins to diagnose CL, which can provide signi cant speci city and sensitivity levels by using a single puri ed antigen (Rezende et al 2016). Efforts to characterize the bacterial proteome and to discover new secreted antigens with potential for use in vaccine development and immunoassays against C. pseudotuberculosis infection are being made (Raynal et al 2018). Thus, the good performance of the ELISA test using rSodC protein demonstrated that it is promising for epidemiological research and CL control programs due to its accuracy in detecting goats and sheep infected with C. pseudotuberculosis.…”

Section: Discussionmentioning

confidence: 99%

rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

Farias¹,

Filho

Marchioro

et al. 2020

Preprint

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Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that recombinant SodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.

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“…Few methods use recombinant proteins to diagnose CL, which can provide significant specificity and sensitivity levels by using a single purified antigen (Rezende et al 2016). Efforts to characterize the bacterial proteome and to discover new secreted antigens with potential for use in vaccine development and immunoassays against C. pseudotuberculosis infection are being made (Raynal et al 2018). Thus, the good performance of the ELISA test using rSodC protein demonstrated that it is promising for epidemiological research and CL control programs due to its accuracy in detecting goats and sheep infected with C. pseudotuberculosis.…”

Section: Discussionmentioning

confidence: 99%

“…Considering that the use of an efficient diagnostic method is crucial for the success of infectious disease control programs (Bastos et al 2012;Rezende et al 2016), and that extracellular or cell surface antigens allow for better recognition in ELISA tests (Oreiby 2015;Raynal et al 2018), the objective of this study was to evaluate C. pseudotuberculosis rDTxR, rTrx, TrxR, rLexA, rNanH, rSodC, rPknG and rSpaC proteins to develop a CL diagnostic test using serum from naturally and experimentally infected goats and sheep.…”

Section: Introductionmentioning

confidence: 99%

rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

et al. 2020

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View full text Add to dashboard Cite

Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs. Trial registration AEC No 4958051018. 12/18/2018, retrospectively registered

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Identification of membrane-associated proteins with pathogenic potential expressed by Corynebacterium pseudotuberculosis grown in animal serum

Cited by 9 publications

References 33 publications

Membrane proteomic analysis reveals overlapping and independent functions of Streptococcus mutans Ffh, YidC1, and YidC2

Membrane proteomic analysis reveals overlapping and independent functions of Streptococcus mutans Ffh, YidC1, and YidC2

rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

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