A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization.
Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. Substitution of MOMP Thr(268) led to significantly reduced binding to BgAgs. The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis.
School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
Jafar
J
Mahdavi
Necmettin
N
Pirinccioglu
Neil J
NJ
Oldfield
Elisabet
E
Carlsohn
Jeroen
J
Stoof
Akhmed
A
Aslam
Tim
T
Self
Shaun A
SA
Cawthraw
Liljana
L
Petrovska
Natalie
N
Colborne
Carina
C
Sihlbom
Thomas
T
Borén
Karl G
KG
Wooldridge
Dlawer A A
DA
Ala'Aldeen
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D018441:Biofilms; D001789:Blood Group Antigens / Q000737:chemistry / Q000378:metabolism*; D018938:Caco-2 Cells; D016123:Campylobacter jejuni / Q000378:metabolism*; D002645:Chickens; D005408:Flagellin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008024:Ligands; D062105:Molecular Docking Simulation; D016296:Mutagenesis; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D018272:Porins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D001426:Bacterial Proteins; D001789:Blood Group Antigens; D008024:Ligands; D011134:Polysaccharides; D018272:Porins; C095080:major outer membrane protein, Campylobacter jejuni; D005408:Flagellin; C067138:flaA protein, bacteria
Campylobacter jejuni; FlaA; O-glycosylation; biofilm; histo-blood group antigens; major outer membrane protein