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The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions

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The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions. / Paes, Wayne; Dowle, Adam; Coldwell, Jamie; Leech, Andrew; Ganderton, Tim; Brzozowski, Andrzej.

In: PLoS ONE, Vol. 13, No. 6, e0198662, 18.06.2018.

Research output: Contribution to journalArticlepeer-review

Harvard

Paes, W, Dowle, A, Coldwell, J, Leech, A, Ganderton, T & Brzozowski, A 2018, 'The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions', PLoS ONE, vol. 13, no. 6, e0198662. https://doi.org/10.1371/journal.pone.0198662

APA

Paes, W., Dowle, A., Coldwell, J., Leech, A., Ganderton, T., & Brzozowski, A. (2018). The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions. PLoS ONE, 13(6), [e0198662]. https://doi.org/10.1371/journal.pone.0198662

Vancouver

Paes W, Dowle A, Coldwell J, Leech A, Ganderton T, Brzozowski A. The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions. PLoS ONE. 2018 Jun 18;13(6). e0198662. https://doi.org/10.1371/journal.pone.0198662

Author

Paes, Wayne ; Dowle, Adam ; Coldwell, Jamie ; Leech, Andrew ; Ganderton, Tim ; Brzozowski, Andrzej. / The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions. In: PLoS ONE. 2018 ; Vol. 13, No. 6.

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@article{758731762351452d8cbc79b52bd7d40d,
title = "The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions",
abstract = "Chlamydia trachomatis (Ct) is the most common sexually transmitted bacterial pathogen, and the leading cause of infectious blindness worldwide. We have recently shown that immunization with the highly conserved antigenic passenger domain of recombinant Ct polymorphic membrane protein D (rPmpD) is protective in the mouse model of Ct genital tract infection, and previously, that ocular anti-rPmpD antibodies are elicited following vaccination. However, the mechanisms governing the assembly and structure-function relationship of PmpD are unknown. Here, we provide a biophysical analysis of this immunogenic 65 kDa passenger domain fragment of PmpD. Using differential cysteine labeling coupled with LC-MS/MS analysis, we show that widespread intra- and intermolecular disulphide interactions play important roles in the preservation of native monomeric secondary structure and the formation of higher-order oligomers. While it has been proposed that FxxN and GGA(I, L,V) repeat motifs in the Pmp21 ortholog in Chlamydia pneumoniae mediate self-interaction, no such role has previously been identified for cysteine residues in chlamydial Pmps. Further characterisation reveals that oligomeric proteoforms and rPmpD monomers adopt β–sheet folds, consistent with previously described Gram-negative bacterial type V secretion systems (T5SSs). We also highlight adhesin-like properties of rPmpD, showing that both soluble rPmpD and anti-rPmpD serum from immunized mice abrogate binding of rPmpD-coated beads to mammalian cells in a dose-dependent fashion. Hence, our study provides further evidence that chlamydial Pmps may function as adhesins, while elucidating yet another important mechanism of self-association of bacterial T5SS virulence factors that may be unique to the Chlamydiaceae.",
author = "Wayne Paes and Adam Dowle and Jamie Coldwell and Andrew Leech and Tim Ganderton and Andrzej Brzozowski",
note = " {\textcopyright} 2018 Paes et al. ",
year = "2018",
month = jun,
day = "18",
doi = "10.1371/journal.pone.0198662",
language = "English",
volume = "13",
journal = "PLoS ONE",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "6",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - The Chlamydia trachomatis PmpD adhesin forms higher order structures through disulphide-mediated covalent interactions

AU - Paes, Wayne

AU - Dowle, Adam

AU - Coldwell, Jamie

AU - Leech, Andrew

AU - Ganderton, Tim

AU - Brzozowski, Andrzej

N1 - © 2018 Paes et al.

PY - 2018/6/18

Y1 - 2018/6/18

N2 - Chlamydia trachomatis (Ct) is the most common sexually transmitted bacterial pathogen, and the leading cause of infectious blindness worldwide. We have recently shown that immunization with the highly conserved antigenic passenger domain of recombinant Ct polymorphic membrane protein D (rPmpD) is protective in the mouse model of Ct genital tract infection, and previously, that ocular anti-rPmpD antibodies are elicited following vaccination. However, the mechanisms governing the assembly and structure-function relationship of PmpD are unknown. Here, we provide a biophysical analysis of this immunogenic 65 kDa passenger domain fragment of PmpD. Using differential cysteine labeling coupled with LC-MS/MS analysis, we show that widespread intra- and intermolecular disulphide interactions play important roles in the preservation of native monomeric secondary structure and the formation of higher-order oligomers. While it has been proposed that FxxN and GGA(I, L,V) repeat motifs in the Pmp21 ortholog in Chlamydia pneumoniae mediate self-interaction, no such role has previously been identified for cysteine residues in chlamydial Pmps. Further characterisation reveals that oligomeric proteoforms and rPmpD monomers adopt β–sheet folds, consistent with previously described Gram-negative bacterial type V secretion systems (T5SSs). We also highlight adhesin-like properties of rPmpD, showing that both soluble rPmpD and anti-rPmpD serum from immunized mice abrogate binding of rPmpD-coated beads to mammalian cells in a dose-dependent fashion. Hence, our study provides further evidence that chlamydial Pmps may function as adhesins, while elucidating yet another important mechanism of self-association of bacterial T5SS virulence factors that may be unique to the Chlamydiaceae.

AB - Chlamydia trachomatis (Ct) is the most common sexually transmitted bacterial pathogen, and the leading cause of infectious blindness worldwide. We have recently shown that immunization with the highly conserved antigenic passenger domain of recombinant Ct polymorphic membrane protein D (rPmpD) is protective in the mouse model of Ct genital tract infection, and previously, that ocular anti-rPmpD antibodies are elicited following vaccination. However, the mechanisms governing the assembly and structure-function relationship of PmpD are unknown. Here, we provide a biophysical analysis of this immunogenic 65 kDa passenger domain fragment of PmpD. Using differential cysteine labeling coupled with LC-MS/MS analysis, we show that widespread intra- and intermolecular disulphide interactions play important roles in the preservation of native monomeric secondary structure and the formation of higher-order oligomers. While it has been proposed that FxxN and GGA(I, L,V) repeat motifs in the Pmp21 ortholog in Chlamydia pneumoniae mediate self-interaction, no such role has previously been identified for cysteine residues in chlamydial Pmps. Further characterisation reveals that oligomeric proteoforms and rPmpD monomers adopt β–sheet folds, consistent with previously described Gram-negative bacterial type V secretion systems (T5SSs). We also highlight adhesin-like properties of rPmpD, showing that both soluble rPmpD and anti-rPmpD serum from immunized mice abrogate binding of rPmpD-coated beads to mammalian cells in a dose-dependent fashion. Hence, our study provides further evidence that chlamydial Pmps may function as adhesins, while elucidating yet another important mechanism of self-association of bacterial T5SS virulence factors that may be unique to the Chlamydiaceae.

UR - http://www.scopus.com/inward/record.url?scp=85048835169&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0198662

DO - 10.1371/journal.pone.0198662

M3 - Article

AN - SCOPUS:85048835169

VL - 13

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 6

M1 - e0198662

ER -