title: P159 from Mycoplasma hyopneumoniae binds porcine cilia and heparin and is cleaved in a manner akin to ectodomain shedding.
abstract: Mycoplasma hyopneumoniae colonizes the ciliated epithelial lining of the upper respiratory tract of swine and results in chronic infection. Previously, we have observed that members of P97 and P102 paralog families of cilium adhesins undergo endoproteolytic processing on the surface of M. hyopneumoniae. We show that P159 (MHJ_0494), an epithelial cell adhesin unrelated to P97 and P102 paralog families, is a cilium adhesin that undergoes dominant cleavage events at S/T-X-F↓X-D/E-like motifs located at positions (233)F↓Q(234) and (981)F↓Q(982), generating P27, P110, and P52. An unrelated cleavage site (738)L-K-V↓G-A-A(743) in P110 shows sequence identity with a cleavage site (L-N-V↓A-V-S) identified in the P97 paralog, Mhp385, and generates 76 (P76) and 35 kDa (P35) fragments. LC-MS/MS analysis of biotinylated surface proteins identified six peptides with a biotin moiety on their N-terminus indicating novel, low abundance neo-N-termini. LC-MS/MS of proteins separated by 2D-PAGE, 2D immunoblotting using monospecific antiserum raised against recombinant fragments spanning P159 (F1(P159)-F4(P159)), and proteins that bound to heparin-agarose were all used to map P159 cleavage fragments. P159 is the first cilium adhesin not belonging to the P97/P102 paralog families and is extensively processed in a manner akin to ectodomain shedding in eukaryotes.
authors.affiliation: The ithree institute, University of Technology Sydney , PO Box 123, Broadway, NSW 2007, Australia.
authors.forename: Benjamin B A
authors.initials: BB
authors.lastname: Raymond
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Tacchi
authors.forename: Veronica M
authors.initials: VM
authors.lastname: Jarocki
authors.forename: F Chris
authors.initials: FC
authors.lastname: Minion
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 / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019921:Biotinylation; D002853:Chromatography, Liquid; D002923:Cilia / Q000737:chemistry* / Q000378:metabolism; D006493:Heparin / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D045705:Mycoplasma hyopneumoniae / Q000737:chemistry* / Q000502:physiology; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D010449:Peptide Mapping; D045729:Pneumonia of Swine, Mycoplasmal / Q000382:microbiology; D011485:Protein Binding; D059748:Proteolysis; D012137:Respiratory System / Q000382:microbiology; D013552:Swine; D053719:Tandem Mass Spectrometry
chemical_list: D018829:Adhesins, Bacterial; D010446:Peptide Fragments; D006493:Heparin