From the Institute for Biological Sciences, National Research Council, Ottawa, Ontario K1A 0R6, Canada
Campylobacter jejuni is well known for synthesizing ganglioside mimics within the glycan component of its lipooligosaccharide (LOS), which have been implicated in triggering Guillain-Barre syndrome. We now confirm that this pathogen is capable of synthesizing a much broader spectrum of host glycolipid/glycoprotein mimics within its LOS. P blood group and paragloboside (lacto-N-neotetraose) antigen mimicry is exhibited by RM1221, a strain isolated from a poultry source. RM1503, a gastroenteritis-associated strain, expresses lacto-N-biose and sialyl-Lewis c units, the latter known as the pancreatic tumor-associated antigen, DU-PAN-2 (or LSTa). C. jejuni GC149, a Guillain-Barre syndrome-associated strain, expresses an unusual sialic acid-containing hybrid oligosaccharide with similarity to both ganglio and P(k) antigens and can, through phase variation of its LOS biosynthesis genes, display GT1a or GD3 ganglioside mimics. We show that the sialyltransferase CstII and the galactosyltransferase CgtD are involved in the synthesis of multiple mimic types, with LOS structural diversity achieved through evolving allelic substrate specificity.
13C NMR, 1H NMR, methylation, NMR-2D, GC-MS, sugar analysis, 31P NMR, NMR-1D, genetic methods, de-N-O-acylation, LC-MS, CE
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6 C7
5,2 bDGlcp 102.5 73.5 76.3 70.1 76.9 61.1
5,3,2 bDGalp 103.9 71.4 73.1 69.3 76.6 62.0
5,3 a?Hepp 101.1 78.2 71.0 67.4 71.9 68.7 63.8
5,4,4,3,4,4,3,2 Ac
5,4,4,3,4,4,3 bDGalpN 104.2 53.5 71.7 68.7 75.9 62.0
5,4,4,3,4,4 aDGalp 101.3 68.6 79.6 69.9 71.2 61.3
5,4,4,3,4 bDGalp 104.2 71.8 73.1 78.1 76.4 61.2
5,4,4,3,2 Ac
5,4,4,3 bDGlcpN 103.6 56.3 73.1 79.5 75.6 60.9
5,4,4 bDGalp 103.6 71.0 82.7 69.5 76.2 62.1
5,4 bDGlcp 103.5 74.2 75.0 79.8 77.0 62.2
5 a?Hepp 98.9 78.0 73.6 74.6 72.3 68.6 64.0
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1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6 H7
5,2 bDGlcp 4.42 3.38 3.51 3.47 3.39 3.78-3.91
5,3,2 bDGalp 4.29 3.57 3.67 3.94 3.56 3.76-3.85
5,3 a?Hepp 5.24 4.23 3.90 3.91 3.89 4.10 3.63-3.74
5,4,4,3,4,4,3,2 Ac
5,4,4,3,4,4,3 bDGalpN 4.65 3.95 3.77 3.95 3.69 3.77-3.81
5,4,4,3,4,4 aDGalp 4.93 3.91 3.97 4.27 4.40 3.71
5,4,4,3,4 bDGalp 4.56 3.60 3.77 4.06 3.80 ?
5,4,4,3,2 Ac
5,4,4,3 bDGlcpN 4.72 3.81 3.74 3.76 3.60 3.89-4.00
5,4,4 bDGalp 4.44 3.60 3.73 4.14 3.64 3.75-3.77
5,4 bDGlcp 4.57 3.36 3.69 3.40 3.66 3.72-4.08
5 a?Hepp 5.21 4.27 4.27 4.20 4.20 4.18 3.74-3.74
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1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6 C7/H7
5,2 bDGlcp 102.5/4.42 73.5/3.38 76.3/3.51 70.1/3.47 76.9/3.39 61.1/3.78-3.91
5,3,2 bDGalp 103.9/4.29 71.4/3.57 73.1/3.67 69.3/3.94 76.6/3.56 62.0/3.76-3.85
5,3 a?Hepp 101.1/5.24 78.2/4.23 71.0/3.90 67.4/3.91 71.9/3.89 68.7/4.10 63.8/3.63-3.74
5,4,4,3,4,4,3,2 Ac
5,4,4,3,4,4,3 bDGalpN 104.2/4.65 53.5/3.95 71.7/3.77 68.7/3.95 75.9/3.69 62.0/3.77-3.81
5,4,4,3,4,4 aDGalp 101.3/4.93 68.6/3.91 79.6/3.97 69.9/4.27 71.2/4.40 61.3/3.71
5,4,4,3,4 bDGalp 104.2/4.56 71.8/3.60 73.1/3.77 78.1/4.06 76.4/3.80 61.2/?
5,4,4,3,2 Ac
5,4,4,3 bDGlcpN 103.6/4.72 56.3/3.81 73.1/3.74 79.5/3.76 75.6/3.60 60.9/3.89-4.00
5,4,4 bDGalp 103.6/4.44 71.0/3.60 82.7/3.73 69.5/4.14 76.2/3.64 62.1/3.75-3.77
5,4 bDGlcp 103.5/4.57 74.2/3.36 75.0/3.69 79.8/3.40 77.0/3.66 62.2/3.72-4.08
5 a?Hepp 98.9/5.21 78.0/4.27 73.6/4.27 74.6/4.20 72.3/4.20 68.6/4.18 64.0/3.74-3.74
?XKdo?
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