Glycoconjugate Journal

, 26:1055 | Cite as

Identification of high-mannose and multiantennary complex-type N-linked glycans containing α-galactose epitopes from Nurse shark IgM heavy chain

  • David J. Harvey
  • Max Crispin
  • Beryl E. Moffatt
  • Sylvia L. Smith
  • Robert B. Sim
  • Pauline M. Rudd
  • Raymond A. Dwek


MALDI-TOF mass spectrometry, negative ion nano-electrospray MS/MS and exoglycosidase digestion were used to identify 36 N-linked glycans from 19S IgM heavy chain derived from the nurse shark (Ginglymostoma cirratum). The major glycan was the high-mannose compound, Man6GlcNAc2 accompanied by small amounts of Man5GlcNAc2, Man7GlcNAc2 and Man8GlcNAc2. Bi- and tri-antennary (isomer with a branched 3-antenna) complex-type glycans were also abundant, most contained a bisecting GlcNAc residue (β1→4-linked to the central mannose) and with varying numbers of α-galactose residues capping the antennae. Small amounts of monosialylated glycans were also found. This appears to be the first comprehensive study of glycosylation in this species of animal. The glycosylation pattern has implications for the mechanism of activation of the complement system by nurse shark IgM.


Nurse shark N-Glycans MALDI mass spectrometry Negative ion fragmentation α-Galactose 



Arthrobacter ureafaciens sialidase


bovine testis β-galactosidase


Green coffee-bean α-galactosidase


dihydroxybenzoic acid




ethylenediamine tetra-acetic acid










immunoglobulin M


matrix-assisted laser desorption/ionization




mannose-binding lectin


mass spectrometry


polyacrylamide gel electrophoresis


protein N-glycosidase




sodium dodecylsulphate


Streptococcus pneumoniae β-N-acetylhexosaminidase





We thank the Wellcome Trust and the Biotechnology and Biological Sciences Research Council for equipment grants to purchase the Q-Tof and TofSpec mass spectrometers respectively. SS was supported by NIH/MBRS grant SO6 GM008205.


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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • David J. Harvey
    • 1
  • Max Crispin
    • 2
  • Beryl E. Moffatt
    • 3
  • Sylvia L. Smith
    • 4
  • Robert B. Sim
    • 3
  • Pauline M. Rudd
    • 5
  • Raymond A. Dwek
    • 1
  1. 1.Oxford Glycobiology Institute, Department of BiochemistryUniversity of OxfordOxfordUK
  2. 2.Division of Structural Biology, Wellcome Trust Centre for Human GeneticsUniversity of OxfordOxfordUK
  3. 3.MRC Immunochemistry Unit, Department of BiochemistryUniversity of OxfordOxfordUK
  4. 4.Department of Biological SciencesFlorida International UniversityMiamiUSA
  5. 5.National Institute for Bioprocessing, Research and Training (NIBRT), Conway InstituteUniversity College DublinDublin 4Ireland

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