title: Proteolytic processing of the cilium adhesin MHJ_0194 (P123J ) in Mycoplasma hyopneumoniae generates a functionally diverse array of cleavage fragments that bind multiple host molecules.
abstract: Mycoplasma hyopneumoniae, the aetiological agent of porcine enzootic pneumonia, regulates the presentation of proteins on its cell surface via endoproteolysis, including those of the cilial adhesin P123 (MHJ_0194). These proteolytic cleavage events create functional adhesins that bind to proteoglycans and glycoproteins on the surface of ciliated and non-ciliated epithelial cells and to the circulatory host molecule plasminogen. Two dominant cleavage events of the P123 preprotein have been previously characterized; however, immunoblotting studies suggest that more complex processing events occur. These extensive processing events are characterized here. The functional significance of the P97 cleavage fragments is also poorly understood. Affinity chromatography using heparin, fibronectin and plasminogen as bait and peptide arrays were used to expand our knowledge of the adhesive capabilities of P123 cleavage fragments and characterize a novel binding motif in the C-terminus of P123. Further, we use immunohistochemistry to examine in vivo, the biological significance of interactions between M. hyopneumoniae and fibronectin and show that M. hyopneumoniae induces fibronectin deposition at the site of infection on the ciliated epithelium. Our data supports the hypothesis that M. hyopneumoniae possesses the molecular machinery to influence key molecular communication pathways in host cells.
authors.affiliation: The ithree Institute, University of Technology, Sydney, NSW, Australia.
authors.forename: Benjamin B A
authors.initials: BB
authors.lastname: Raymond
authors.forename: Cheryl
authors.initials: C
authors.lastname: Jenkins
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Seymour
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Tacchi
authors.forename: Michael
authors.initials: M
authors.lastname: Widjaja
authors.forename: Veronica M
authors.initials: VM
authors.lastname: Jarocki
authors.forename: Ania T
authors.initials: AT
authors.lastname: Deutscher
authors.forename: Lynne
authors.initials: L
authors.lastname: Turnbull
authors.forename: Cynthia B
authors.initials: CB
authors.lastname: Whitchurch
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Padula
authors.forename: Steven P
authors.initials: SP
authors.lastname: Djordjevic
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D005353:Fibronectins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D045705:Mycoplasma hyopneumoniae / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D040081:Protein Array Analysis; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D053719:Tandem Mass Spectrometry
chemical_list: D018829:Adhesins, Bacterial; D005353:Fibronectins; D006023:Glycoproteins; D011134:Polysaccharides