Mhp493 (P216) is a proteolytically processed, cilium and heparin binding protein of Mycoplasma hyopneumoniade

Jody Wilton, Cheryl Jenkins, Stuart J. Cordwell, Linda Falconer, F. Chris Minion, David C. Oneal, Michael A. Djordjevic, Angela Connolly, Idris Barchia, Mark J. Walker, Steven P. Djordjevic

    Research output: Contribution to journalArticlepeer-review

    60 Citations (Scopus)

    Abstract

    Mycoplasma hyopneumoniae induces respiratory disease in swine by colonizing cilia causing ciliostasis, cilial loss and epithelial cell death. Heparin binds to M. hyopneumoniae cells in a dose-dependent manner and blocks its ability to adhere to porcine cilia. We show here that Mhp493 (P216), a paralogue of the cilium adhesin P97 (Mhp183), is cleaved between amino acids 1040 and 1089 generating surface-accessible, heparin-binding proteins P120 and P85. Antiphosphoserine antibodies recognized P85 in 2-D immunoblotting studies and TiO2 chromatography of trypsin digests of P85 isolated a single peptide with an m/z of 917.3. A phosphoserine residue in the tryptic peptide 90VSELpSFR96 (position 94 in P85) was identified by MALDI-MS/MS. Polyhistidine fusion proteins (F1P216, F2 P216, F3P216) spanning Mhp493 bound heparin with biologically significant Kd values, and heparin, fucoidan and mucin inhibited this interaction. Latex beads coated with F1P216, F2P216 and F3P216 adhered to and entered porcine kidney epithelial-like (PK15) cell monolayers. Microtitre plate-based assays showed that sequences within P120 and P85 bind to porcine cilia and are recognized by serum antibodies elicited during infection by M. hyopneumoniae. Mhp493 contributes significantly to the surface architecture of M. hyopneumoniae and is the first cilium adhesin to be described that lacks an R1 cilium-binding domain.

    Original languageEnglish
    Pages (from-to)566-582
    Number of pages17
    JournalMolecular Microbiology
    Volume71
    Issue number3
    DOIs
    Publication statusPublished - Feb 2009

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