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

Glycoconjugate Journal volume 26, Article number: 1055 (2009) Cite this article

Abstract

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.

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Abbreviations

ABS:

Arthrobacter ureafaciens sialidase

BTG:

bovine testis β-galactosidase

CBG:

Green coffee-bean α-galactosidase

DHB:

dihydroxybenzoic acid

dHex:

deoxy-hexose

EDTA:

ethylenediamine tetra-acetic acid

Gal:

galactose

GlcNAc:

N-acetylglucosamine

Hex:

hexose

HexNAc:

N-acetylhexosamine

IgM:

immunoglobulin M

MALDI:

matrix-assisted laser desorption/ionization

Man:

mannose

MBL:

mannose-binding lectin

MS:

mass spectrometry

PAGE:

polyacrylamide gel electrophoresis

PNGase:

protein N-glycosidase

Q:

quadrupole

SDS:

sodium dodecylsulphate

SPH:

Streptococcus pneumoniae β-N-acetylhexosaminidase

TOF:

time-of-flight

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Acknowledgements

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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Affiliations

  1. Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK

    David J. Harvey & Raymond A. Dwek

  2. Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7BN, UK

    Max Crispin

  3. MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK

    Beryl E. Moffatt & Robert B. Sim

  4. Department of Biological Sciences, Florida International University, Miami, FL, 33199, USA

    Sylvia L. Smith

  5. National Institute for Bioprocessing, Research and Training (NIBRT), Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland

    Pauline M. Rudd

Authors
  1. David J. Harvey
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  2. Max Crispin
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  3. Beryl E. Moffatt
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  4. Sylvia L. Smith
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  5. Robert B. Sim
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  6. Pauline M. Rudd
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  7. Raymond A. Dwek
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Correspondence to David J. Harvey.

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Harvey, D.J., Crispin, M., Moffatt, B.E. et al. Identification of high-mannose and multiantennary complex-type N-linked glycans containing α-galactose epitopes from Nurse shark IgM heavy chain. Glycoconj J 26, 1055 (2009). https://doi.org/10.1007/s10719-008-9226-5

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  • DOI: https://doi.org/10.1007/s10719-008-9226-5

Keywords

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