The N-linking glycosylation system from Actinobacillus pleuropneumoniae is required for adhesion and has potential use in glycoengineering

File Description SizeFormat 
160212.full.pdfPublished version1.18 MBAdobe PDFView/Open
Title: The N-linking glycosylation system from Actinobacillus pleuropneumoniae is required for adhesion and has potential use in glycoengineering
Authors: Cuccui, J
Terra, VS
Bossé, JT
Naegeli, A
Abouelhadid, S
Li, Y
Lin, CW
Vohra, P
Tucker, AW
Rycroft, AN
Maskell, DJ
Aebi, M
Langford, PR
Wren, BW
BRaDP1T Consortium
Item Type: Journal Article
Abstract: Actinobacillus pleuropneumoniae is a mucosal respiratory pathogen causing contagious porcine pleuropneumonia. Pathogenesis studies have demonstrated a major role for the capsule, exotoxins and outer membrane proteins. Actinobacillus pleuropneumoniae can also glycosylate proteins, using a cytoplasmic N-linked glycosylating enzyme designated NGT, but its transcriptional arrangement and role in virulence remains unknown. We investigated the NGT locus and demonstrated that the putative transcriptional unit consists of rimO, ngt and a glycosyltransferase termed agt. From this information we used the A. pleuropneumoniae glycosylation locus to decorate an acceptor protein, within Escherichia coli, with a hexose polymer that reacted with an anti-dextran antibody. Mass spectrometry analysis of a truncated protein revealed that this operon could add up to 29 repeat units to the appropriate sequon. We demonstrated the importance of NGT in virulence, by creating deletion mutants and testing them in a novel respiratory cell line adhesion model. This study demonstrates the importance of the NGT glycosylation system for pathogenesis and its potential biotechnological application for glycoengineering.
Issue Date: 11-Jan-2017
Date of Acceptance: 28-Nov-2016
URI: http://hdl.handle.net/10044/1/44223
DOI: http://dx.doi.org/10.1098/rsob.160212
ISSN: 2046-2441
Publisher: Royal Society, The
Journal / Book Title: Open Biology
Volume: 7
Copyright Statement: © 2017 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Pfizer Limited (UK)
Funder's Grant Number: BB/G018553/1
N/A
Keywords: Actinobacillus pleuropneumoniae
N-linked glycosylation
adhesion
Publication Status: Published
Conference Place: England
Open Access location: http://rsob.royalsocietypublishing.org/content/7/1/160212
Article Number: 160212
Appears in Collections:Department of Medicine



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Creative Commonsx