O-linked glycosylation ensures the normal conformation of the autotransporter adhesin involved in diffuse adherence

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DOIResolve DOI: http://doi.org/10.1128/JB.00969-07
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TypeArticle
Journal titleJournal of Bacteriology
Volume189
Issue24
Pages88808889; # of pages: 10
SubjectCells; Epithelial Cells; Escherichia coli; Glycosylation; Heat; In Vitro; Peptides; pha; Protease; Protein; Proteins; Stress; Vitro
AbstractThe Escherichia coli adhesin involved in diffuse adherence (AIDA-I) is one of the few glycosylated proteins found in Escherichia coli. Glycosylation is mediated by a specific heptosyltransferase encoded by the aah gene, but little is known about the role of this modification and the mechanism involved. In this study, we identified several peptides of AIDA-I modified by the addition of heptoses by use of mass spectrometry and N-terminal sequencing of proteolytic fragments of AIDA-I. One threonine and 15 serine residues were identified as bearing heptoses, thus demonstrating for the first time that AIDA-I is O-glycosylated. We observed that unglycosylated AIDA-I is expressed in smaller amounts than its glycosylated counterpart and shows extensive signs of degradation upon heat extraction. We also observed that unglycosylated AIDA-I is more sensitive to proteases and induces important extracytoplasmic stress. Lastly, as was previously shown, we noted that glycosylation is required for AIDA-I to mediate adhesion to cultured epithelial cells, but purified mature AIDA-I fused to GST was found to bind in vitro to cells whether or not it was glycosylated. Taken together, our results suggest that glycosylation is required to ensure a normal conformation of AIDA-I and may be only indirectly necessary for its cell-binding function
Publication date
LanguageEnglish
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedNo
NRC number47555
NPARC number3539460
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Record identifier2bcabfd2-eb9c-454a-90d9-d4bec509883e
Record created2009-03-01
Record modified2016-05-09
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