Cilium Adhesin P216 (MHJ-0493) is a target of ectodomain shedding and aminopeptidase activity on the surface of mycoplasma hyopneumoniae

Tacchi, JL; Raymond, BBA; Jarocki, VM; Berry, IJ; Padula, MP; Djordjevic, SP

Cilium Adhesin P216 (MHJ-0493) is a target of ectodomain shedding and aminopeptidase activity on the surface of mycoplasma hyopneumoniae

Tacchi, JL Raymond, BBA

Jarocki, VM

Berry, IJ

Padula, MP

Djordjevic, SP

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Publication Type:: Journal Article
Citation:: Journal of Proteome Research, 2014, 13 (6), pp. 2920 - 2930
Issue Date:: 2014-06-06

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Field	Value	Language
dc.contributor.author	Tacchi, JL	en_US
dc.contributor.author	Raymond, BBA https://orcid.org/0000-0003-3610-9289	en_US
dc.contributor.author	Jarocki, VM https://orcid.org/0000-0003-1249-0994	en_US
dc.contributor.author	Berry, IJ https://orcid.org/0000-0002-0322-2595	en_US
dc.contributor.author	Padula, MP https://orcid.org/0000-0002-8283-0643	en_US
dc.contributor.author	Djordjevic, SP https://orcid.org/0000-0001-9301-5372	en_US
dc.date.issued	2014-06-06	en_US
dc.identifier.citation	Journal of Proteome Research, 2014, 13 (6), pp. 2920 - 2930	en_US
dc.identifier.issn	1535-3893	en_US
dc.identifier.uri	http://hdl.handle.net/10453/37994
dc.description.abstract	MHJ-0493 (P216) is a highly expressed cilium adhesin in Mycoplasma hyopneumoniae. P216 undergoes cleavage at position 1074 in the S/T-X-F↓-X-D/E-like motif 1072T-N-F↓Q-E1076 generating N-terminal and C-terminal fragments of 120 kDa (P120) and 85 kDa (P85) on the surface of M. hyopneumoniae. Here we show that several S/T-X-F↓X-D/E-like motifs exist in P216 but only 1072T-N- F↓Q-E1076 and 1344I-T-F↓A-D-Y1349 were determined to be bona fide processing sites by identifying semitryptic peptides consistent with cleavage at the phenylalanine residue. The location of S/T-X-F↓-X-D/E-like motifs within or abutting regions of protein disorder greater than 40 consecutive amino acids is consistent with our hypothesis that site access influences the cleavage efficiency. Approximately 20 cleavage fragments of P216 were identified on the surface of M. hyopneumoniae by LC-MS/MS analysis of biotinylated proteins and 2D SDS-PAGE. LC-MS/MS analysis of semitryptic peptides within P216 identified novel cleavage sites. Moreover, detection of a series of overlapping semitryptic peptides that differed by the loss a single amino acid at their N-terminus is consistent with aminopeptidase activity on the surface of M. hyopneumoniae. P120 and P85 and their cleavage fragments bind heparin and cell-surface proteins derived from porcine epithelial-like cells, indicating that P216 cleavage fragments retain the ability to bind glycosaminoglycans. © 2014 American Chemical Society.	en_US
dc.relation.ispartof	Journal of Proteome Research	en_US
dc.relation.isbasedon	10.1021/pr500087c	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Cell Line	en_US
dc.subject.mesh	Epithelial Cells	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Sus scrofa	en_US
dc.subject.mesh	Mycoplasma hyopneumoniae	en_US
dc.subject.mesh	Aminopeptidases	en_US
dc.subject.mesh	Heparin	en_US
dc.subject.mesh	Peptide Fragments	en_US
dc.subject.mesh	Adhesins, Bacterial	en_US
dc.subject.mesh	Membrane Proteins	en_US
dc.subject.mesh	Peptide Mapping	en_US
dc.subject.mesh	Bacterial Adhesion	en_US
dc.subject.mesh	Binding Sites	en_US
dc.subject.mesh	Amino Acid Motifs	en_US
dc.subject.mesh	Protein Binding	en_US
dc.subject.mesh	Molecular Sequence Data	en_US
dc.subject.mesh	Tandem Mass Spectrometry	en_US
dc.subject.mesh	Proteolysis	en_US
dc.title	Cilium Adhesin P216 (MHJ-0493) is a target of ectodomain shedding and aminopeptidase activity on the surface of mycoplasma hyopneumoniae	en_US
dc.type	Journal Article
utslib.citation.volume	6	en_US
utslib.citation.volume	13	en_US
utslib.for	060109 Proteomics and Intermolecular Interactions (excl. Medical Proteomics)	en_US
utslib.for	060106 Cellular Interactions (incl. Adhesion, Matrix, Cell Wall)	en_US
utslib.for	03 Chemical Sciences	en_US
utslib.for	06 Biological 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	6	en_US
pubs.publication-status	Published	en_US
pubs.volume	13	en_US

Abstract:

MHJ-0493 (P216) is a highly expressed cilium adhesin in Mycoplasma hyopneumoniae. P216 undergoes cleavage at position 1074 in the S/T-X-F↓-X-D/E-like motif 1072T-N-F↓Q-E1076 generating N-terminal and C-terminal fragments of 120 kDa (P120) and 85 kDa (P85) on the surface of M. hyopneumoniae. Here we show that several S/T-X-F↓X-D/E-like motifs exist in P216 but only 1072T-N- F↓Q-E1076 and 1344I-T-F↓A-D-Y1349 were determined to be bona fide processing sites by identifying semitryptic peptides consistent with cleavage at the phenylalanine residue. The location of S/T-X-F↓-X-D/E-like motifs within or abutting regions of protein disorder greater than 40 consecutive amino acids is consistent with our hypothesis that site access influences the cleavage efficiency. Approximately 20 cleavage fragments of P216 were identified on the surface of M. hyopneumoniae by LC-MS/MS analysis of biotinylated proteins and 2D SDS-PAGE. LC-MS/MS analysis of semitryptic peptides within P216 identified novel cleavage sites. Moreover, detection of a series of overlapping semitryptic peptides that differed by the loss a single amino acid at their N-terminus is consistent with aminopeptidase activity on the surface of M. hyopneumoniae. P120 and P85 and their cleavage fragments bind heparin and cell-surface proteins derived from porcine epithelial-like cells, indicating that P216 cleavage fragments retain the ability to bind glycosaminoglycans. © 2014 American Chemical Society.

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http://hdl.handle.net/10453/37994