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

Wilton, J; Jenkins, C; Cordwell, SJ; Falconer, L; Minion, FC; Oneal, DC; Djordjevic, MA; Connolly, A; Barchia, I; Walker, MJ; Djordjevic, SP

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

Wilton, J Jenkins, C

Cordwell, SJ Falconer, L Minion, FC Oneal, DC Djordjevic, MA Connolly, A Barchia, I Walker, MJ Djordjevic, SP

Permalink

Publication Type:: Journal Article
Citation:: Molecular Microbiology, 2009, 71 (3), pp. 566 - 582
Issue Date:: 2009-02-01

Closed Access

	Filename	Description	Size
	2008007081OK.pdf		798.11 kB	Adobe PDF	View/Open

Copyright Clearance Process

Recently Added
In Progress
Closed Access

This item is closed access and not available.

Full metadata record

Field	Value	Language
dc.contributor.author	Wilton, J	en_US
dc.contributor.author	Jenkins, C https://orcid.org/0000-0002-5966-072X	en_US
dc.contributor.author	Cordwell, SJ	en_US
dc.contributor.author	Falconer, L	en_US
dc.contributor.author	Minion, FC	en_US
dc.contributor.author	Oneal, DC	en_US
dc.contributor.author	Djordjevic, MA	en_US
dc.contributor.author	Connolly, A	en_US
dc.contributor.author	Barchia, I	en_US
dc.contributor.author	Walker, MJ	en_US
dc.contributor.author	Djordjevic, SP https://orcid.org/0000-0001-9301-5372	en_US
dc.date.issued	2009-02-01	en_US
dc.identifier.citation	Molecular Microbiology, 2009, 71 (3), pp. 566 - 582	en_US
dc.identifier.issn	0950-382X	en_US
dc.identifier.uri	http://hdl.handle.net/10453/15027
dc.description.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. © 2008 Blackwell Publishing Ltd.	en_US
dc.relation.ispartof	Molecular Microbiology	en_US
dc.relation.isbasedon	10.1111/j.1365-2958.2008.06546.x	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Cells, Cultured	en_US
dc.subject.mesh	Cilia	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Swine	en_US
dc.subject.mesh	Mycoplasma hyopneumoniae	en_US
dc.subject.mesh	Heparin	en_US
dc.subject.mesh	Adhesins, Bacterial	en_US
dc.subject.mesh	Recombinant Proteins	en_US
dc.subject.mesh	Antibodies, Bacterial	en_US
dc.subject.mesh	Cloning, Molecular	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Protein Binding	en_US
dc.subject.mesh	Genes, Bacterial	en_US
dc.subject.mesh	Molecular Sequence Data	en_US
dc.title	Mhp493 (P216) is a proteolytically processed, cilium and heparin binding protein of Mycoplasma hyopneumoniade	en_US
dc.type	Journal Article
utslib.citation.volume	3	en_US
utslib.citation.volume	71	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	07 Agricultural and Veterinary Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.issue	3	en_US
pubs.publication-status	Published	en_US
pubs.volume	71	en_US

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. © 2008 Blackwell Publishing Ltd.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/15027