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. 2013 Jan 21;8(1):e53999. doi: 10.1371/journal.pone.0053999

Figure 2. Characterization of the El Tor binding glycosphingolipid of human small intestine.

Figure 2

(A) Chemical detection by anisaldehyde. (B) Autoradiogram obtained by binding of V. cholerae strain JBK 70. (C) Autoradiogram obtained by binding of H. pylori strain J99. The lanes were: Lane 1, fraction HI-I from human small intestine, 4 µg; Lane 2, NeuAcα3Galβ4(Fucα3)GlcNAcβ3Galβ4GlcNAcβ3Galβ4Glcβ1Cer (sialyl-Lex-octaosylceramide), 4 µg; Lane 3, Galβ4(Fucα3)GlcNAcβ3Galβ4GlcNAcβ3Galβ4Glcβ1Cer (Lex-heptaosylceramide), 4 µg; Lane 4, NeuAcα3Galβ4GlcNAcβ3Galβ4(Fucα3)GlcNAcβ3Galβ4Glcβ1Cer (VIM2), 4 µg; Lane 5, Galβ4GlcNAcβ3Galβ4(Fucα3)GlcNAcβ3Galβ4Glcβ1Cer (de-sialo VIM2), 4 µg; Lane 6, NeuAcα3Galβ4(Fucα3)GlcNAcβ3Galβ4(Fucα3)GlcNAcβ3Galβ4Glcβ1Cer (sialyl-dimeric-Lex), 4 µg; Lane 7, Galβ4(Fucα3)GlcNAcβ3Galβ4(Fucα3)GlcNAcβ3Galβ4Glcβ1Cer (dimeric-Lex), 4 µg. The sialic acid binding H. pylori strain J99 (C) was used as a control for removal of sialic acid from the compounds in lanes 3, 5 and 7 [14]. (D) Base peak chromatogram from LC-ESI/MS of the oligosaccharides derived from fraction HI-I by hydrolysis with Rhodococcus endoglycoceramidase II. (E) MS2 spectrum of the [M-H+]− ion at m/z 1217 (retention time 22.9 min). (F) Anomeric region of the 600 MHz 1H NMR spectrum of fraction HI-I (30°C). The designations G and H refer to Table 1.