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. 2012 Jun 25;287(33):27851–27862. doi: 10.1074/jbc.M112.376525

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Electrospray mass spectrometry of intact polar flagellin protein from Aeromonas hydrophila AH-3. a, electrospray mass spectrum of intact polar flagellin protein showing a complex envelope of multiply charged protein ions. b, the reconstructed molecular mass profile of the polar flagellin is not as well resolved as the lateral flagellin due to the increased complexity, but does clearly show masses at 35,019, 36,685, 38,376, 40,149, and 41,712 Da. This seems to correspond to FlaB, modified with two, three, four, five, and six heptasaccharide glycan chains, ranging in mass from 1563 to 1773 Da. Similar glycan heterogeneity is also observed in the nLC-MS/MS analysis of tryptic digests. c, feCID of polar flagellin, generated by increasing the cone voltage of the QTOF2 mass spectrometer to 85 V. Clear glycan related oxonium ions are present in the low m/z region, resulting from nonspecific fragmentation of the intact flagellin glycoprotein. , 376-Da sugar; ■ = HexNAc.

Inline graphic — Electrospray mass spectrometry of intact polar flagellin protein from Aeromonas hydrophila AH-3. a, electrospray mass spectrum of intact polar flagellin protein showing a complex envelope of multiply charged protein ions. b, the reconstructed molecular mass profile of the polar flagellin is not as well resolved as the lateral flagellin due to the increased complexity, but does clearly show masses at 35,019, 36,685, 38,376, 40,149, and 41,712 Da. This seems to correspond to FlaB, modified with two, three, four, five, and six heptasaccharide glycan chains, ranging in mass from 1563 to 1773 Da. Similar glycan heterogeneity is also observed in the nLC-MS/MS analysis of tryptic digests. c, feCID of polar flagellin, generated by increasing the cone voltage of the QTOF2 mass spectrometer to 85 V. Clear glycan related oxonium ions are present in the low m/z region, resulting from nonspecific fragmentation of the intact flagellin glycoprotein. , 376-Da sugar; ■ = HexNAc.