Structural characterization of sialylated glycoforms of H. influenzae by electrospray mass spectrometry: fragmentation of protonated and sodiated O-deacylated lipopolysaccharides: Rapid Commun.Mass Spectrom.

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TypeArticle
Journal titleRapid Commun.Mass Spectrom.
Volume21
Issue6
Pages952960; # of pages: 9
SubjectAcylation; bacterial; Canada; chemistry; Electrophoresis,Capillary; Electrospray; EXPRESSION; Genome; Haemophilus; Haemophilus influenzae; HAEMOPHILUS-INFLUENZAE; Ions; LACTOSE; linear; LIPOPOLYSACCHARIDE; Lipopolysaccharides; LOCATION; LPS; mass spectrometry; MASS-SPECTROMETRY; method; Methods; MOLECULAR; MOLECULE; oligosaccharide; Protons; SEQUENCE; SEQUENCES; SPECTRA; Spectrometry,Mass,Electrospray Ionization; STRAIN; STRUCTURAL; STRUCTURAL CHARACTERIZATION; structure; Tandem Mass Spectrometry; UNIT
AbstractSialylated lipopolysaccharide (LPS) glycoforms from Haemophilus influenzae were characterized by tandem mass spectrometry using a new generation hyphenated mass spectrometer which combines a triple quadrupole and a linear ion trap (Q-Trap). The fragmentation of both protonated and sodiated molecular ions from O-deacylated LPS (LPS-OH) obtained in MS(2) experiments in the positive mode was studied. The MS(2) spectra of protonated ions provided unambiguous evidence for the presence and sequence of sialylated lactosamine present in lacto-N-neotetraose oligosaccharide extensions but not for sialyl-lactose structures whilst fragmentation of sodiated adducts, [M+Na](+), afforded information diagnostic of mono- and disialylated lactose extensions. To study this we used a highly sialylated LPS from a H. influenzae strain capable of sialyl-lactose expression only. We then applied the method to the H. influenzae genome strain, Rd, in which glycoforms containing both sialyl-lactose and sialyl-lacto-N-neotetraose were detected from diagnostic B-ions at m/z 638.2 ([Neu5Ac(1) Hex(2)+Na](+)) and 657.2 ([Neu5Ac(1) Hex(1) HexNAc(1)+H](+)). Unique fragmentation patterns provided the locations and sequences of these oligosaccharide extensions. This is the first time both sialylated lactose and sialylated lacto-N-neotetraose units have been detected and characterized by tandem mass spectrometry in the same molecule. This methodology is of general applicability for determination of common sialylated oligosaccharide extension in bacterial LPS
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberLI2007
NPARC number9365008
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Record identifier1526b5f8-6186-4320-a2b8-2e374dbe8c61
Record created2009-07-10
Record modified2016-05-09
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