Sequence TTKF ↓ QE defines the site of proteolytic cleavage in Mhp683 protein, a novel glycosaminoglycan and cilium adhesin of Mycoplasma hyopneumoniae

Bogema, DR; Scott, NE; Padula, MP; Tacchi, JL; Raymond, BBA; Jenkins, C; Cordwell, SJ; Minion, FC; Walker, MJ; Djordjevic, SP

Sequence TTKF ↓ QE defines the site of proteolytic cleavage in Mhp683 protein, a novel glycosaminoglycan and cilium adhesin of Mycoplasma hyopneumoniae

Bogema, DR

Scott, NE Padula, MP

Tacchi, JL Raymond, BBA

Jenkins, C

Cordwell, SJ Minion, FC Walker, MJ Djordjevic, SP

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Publication Type:: Journal Article
Citation:: Journal of Biological Chemistry, 2011, 286 (48), pp. 41217 - 41229
Issue Date:: 2011-12-02

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Field	Value	Language
dc.contributor.author	Bogema, DR https://orcid.org/0000-0001-7013-7326	en_US
dc.contributor.author	Scott, NE	en_US
dc.contributor.author	Padula, MP https://orcid.org/0000-0002-8283-0643	en_US
dc.contributor.author	Tacchi, JL	en_US
dc.contributor.author	Raymond, BBA https://orcid.org/0000-0003-3610-9289	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	Minion, FC	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	2011-12-02	en_US
dc.identifier.citation	Journal of Biological Chemistry, 2011, 286 (48), pp. 41217 - 41229	en_US
dc.identifier.issn	0021-9258	en_US
dc.identifier.uri	http://hdl.handle.net/10453/18178
dc.description.abstract	Mycoplasma hyopneumoniae colonizes the ciliated respiratory epithelium of swine, disrupting mucociliary function and inducing chronic inflammation. P97 and P102 family members are major surface proteins of M. hyopneumoniae and play key roles in colonizing cilia via interactions with glycosaminoglycans and mucin. The p102 paralog, mhp683, and homologs in strains from different geographic origins encode a 135-kDa preprotein (P135) that is cleaved into three fragments identified here as P45 683, P48 683, and P50 683. A peptide sequence (TTKF ↓ QE) was identified surrounding both cleavage sites in Mhp683. N-terminal sequences of P48 683 and P50 683, determined by Edman degradation and mass spectrometry, confirmed cleavage after the phenylalanine residue. A similar proteolytic cleavage site was identified by mass spectrometry in another paralog of the P97/P102 family. Trypsin digestion and surface biotinylation studies showed that P45 683, P48 683, and P50 683 reside on the M. hyopneumoniae cell surface. Binding assays of recombinant proteins F1 683-F5 683, spanning Mhp683, showed saturable and dose-dependent binding to biotinylated heparin that was inhibited by unlabeled heparin, fucoidan, and mucin. F1 683-F5 683 also bound porcine epithelial cilia, and antisera to F2 683 and F5 683 significantly inhibited cilium binding by M. hyopneumoniae cells. These data suggest that P45 683, P48 683, and P50 683 each display cilium- and proteoglycan-binding sites. Mhp683 is the first characterized glycosaminoglycan-binding member of the P102 family. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.	en_US
dc.relation.ispartof	Journal of Biological Chemistry	en_US
dc.relation.isbasedon	10.1074/jbc.M111.226084	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Respiratory Mucosa	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	Glycosaminoglycans	en_US
dc.subject.mesh	Adhesins, Bacterial	en_US
dc.subject.mesh	Bacterial Adhesion	en_US
dc.subject.mesh	Amino Acid Motifs	en_US
dc.title	Sequence TTKF ↓ QE defines the site of proteolytic cleavage in Mhp683 protein, a novel glycosaminoglycan and cilium adhesin of Mycoplasma hyopneumoniae	en_US
dc.type	Journal Article
utslib.citation.volume	48	en_US
utslib.citation.volume	286	en_US
utslib.for	0707 Veterinary Sciences	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	03 Chemical Sciences	en_US
utslib.for	06 Biological 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/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.issue	48	en_US
pubs.publication-status	Published	en_US
pubs.volume	286	en_US

Abstract:

Mycoplasma hyopneumoniae colonizes the ciliated respiratory epithelium of swine, disrupting mucociliary function and inducing chronic inflammation. P97 and P102 family members are major surface proteins of M. hyopneumoniae and play key roles in colonizing cilia via interactions with glycosaminoglycans and mucin. The p102 paralog, mhp683, and homologs in strains from different geographic origins encode a 135-kDa preprotein (P135) that is cleaved into three fragments identified here as P45 683, P48 683, and P50 683. A peptide sequence (TTKF ↓ QE) was identified surrounding both cleavage sites in Mhp683. N-terminal sequences of P48 683 and P50 683, determined by Edman degradation and mass spectrometry, confirmed cleavage after the phenylalanine residue. A similar proteolytic cleavage site was identified by mass spectrometry in another paralog of the P97/P102 family. Trypsin digestion and surface biotinylation studies showed that P45 683, P48 683, and P50 683 reside on the M. hyopneumoniae cell surface. Binding assays of recombinant proteins F1 683-F5 683, spanning Mhp683, showed saturable and dose-dependent binding to biotinylated heparin that was inhibited by unlabeled heparin, fucoidan, and mucin. F1 683-F5 683 also bound porcine epithelial cilia, and antisera to F2 683 and F5 683 significantly inhibited cilium binding by M. hyopneumoniae cells. These data suggest that P45 683, P48 683, and P50 683 each display cilium- and proteoglycan-binding sites. Mhp683 is the first characterized glycosaminoglycan-binding member of the P102 family. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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http://hdl.handle.net/10453/18178