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Biogerontology

, 9:351 | Cite as

N-Glycan profiling in the study of human aging

  • Valerie Vanhooren
  • Wouter Laroy
  • Claude Libert
  • Cuiying ChenEmail author
Methods

Abstract

Most secreted proteins produced by the human body are modified by glycosylation. It is well known that the oligosaccharides (glycans) of glycoproteins are important for initiation of various cellular recognition signals that are essential for the maintenance of the ordered social life of each cell within a multi-cellular organism. The sugar chains can be altered by the physiological or pathophysiological condition of the cell. We describe a detailed protocol for the analysis of N-linked glycans in blood via DNA sequencing equipment-Fluorophore Assisted Carbohydrate Electrophoresis (DSA-FACE). The key features of this technique are its robustness, high throughput, high sensitivity and reliable quantification. Based on DSA-FACE technology, we previously reported that N-glycan profiling of the human serum shows substantial changes with increasing age in three major N-glycan structures. We proposed that measurement of the N-glycan level changes could provide a surrogate marker for general health or for age-related disease progression, and for monitoring the improvement of health after therapy.

Keywords

Glycosylation N-glycan Aging Biomarker DSA-FACE 

Abbreviations

DSA-FACE

DNA Sequencer Assisted (DSA)-Fluorophore-Assisted Carbohydrate Electrophoresis (FACE)

PNGase F

Peptide N-Glycosidase F

APTS

8-Amino-1,3,6-PyreneTriSulfonic acid

GlcNAc

N-AcetylGlucosamine

CE

Capillary Electrophoresis

MS

Mass spectrometry

HPLC

High-performance liquid chromatography

HPAEC-PAD

High-performance anion exchange chromatography with pulsed amperometric detection

NG0A2F

Agalactosylated, core-α-1,6-fucosylated biantennary glycan

NG0A2FB

Agalactosylated, core-α-1,6-fucosylated bisecting biantennary glycan

NA2F

Bigalactosylated, core-α-1,6-fucosylated biantennary glycan

Notes

Acknowledgements

We thank Dr. Amin Bredan for editing the manuscript.

References

  1. 1.
    Varki A (1999) Essentials of glycobiology. Cold spring Harbor Laboratory PressGoogle Scholar
  2. 2.
    Rudd PM, Dwek RA (1997) Rapid, sensitive sequencing of oligosaccharides from glycoproteins. Curr Opin Biotechnol 8:488–497PubMedCrossRefGoogle Scholar
  3. 3.
    Guile GR, Rudd PM, Wing DR, Prime SB, Dwek RA (1996) A rapid high-resolution high-performance liquid chromatographic method for separating glycan mixtures and analyzing oligosaccharide profiles. Anal Biochem 240:210–226PubMedCrossRefGoogle Scholar
  4. 4.
    Suzuki S, Honda S (1998) A tabulated review of capillary electrophoresis of carbohydrates. Electrophoresis 19:2539–2560PubMedCrossRefGoogle Scholar
  5. 5.
    Kuster B, Wheeler SF, Hunter AP, Dwek RA, Harvey DJ (1997) Sequencing of n-linked oligosaccharides directly from protein gels: In-gel deglycosylation followed by matrix-assisted laser desorption/ionization mass spectrometry and normal-phase high-performance liquid chromatography. Anal Biochem 250:82–101PubMedCrossRefGoogle Scholar
  6. 6.
    Callewaert N, Geysens S, Molemans F, Contreras R (2001) Ultrasensitive profiling and sequencing of n-linked oligosaccharides using standard DNA-sequencing equipment. Glycobiology 11:275–281PubMedCrossRefGoogle Scholar
  7. 7.
    Laroy W, Contreras R, Callewaert N (2006) Glycome mapping on DNA sequencing equipment. Nat Protoc 1:397–405PubMedCrossRefGoogle Scholar
  8. 8.
    Kukuruzinska MA, Lennon K (1998) Protein n-glycosylation: molecular genetics and functional significance. Crit Rev Oral Biol Med 9:415–448PubMedCrossRefGoogle Scholar
  9. 9.
    Raman R, Raguram S, Venkataraman G, Paulson JC, Sasisekharan R (2005) Glycomics: an integrated systems approach to structure-function relationships of glycans. Nat Methods 2:817–824PubMedCrossRefGoogle Scholar
  10. 10.
    Field MC, Amatayakul-Chantler S, Rademacher TW, Rudd PM, Dwek RA (1994) Structural analysis of the n-glycans from human immunoglobulin a1: Comparison of normal human serum immunoglobulin a1 with that isolated from patients with rheumatoid arthritis. Biochem J 299(Pt 1):261–275PubMedGoogle Scholar
  11. 11.
    Martin K, Talukder R, Hay FC, Axford JS (2001) Characterization of changes in igg associated oligosaccharide profiles in rheumatoid arthritis, psoriatic arthritis, and ankylosing spondylitis using fluorophore linked carbohydrate electrophoresis. J Rheumatol 28:1531–1536PubMedGoogle Scholar
  12. 12.
    Matei L (1997) Plasma proteins glycosylation and its alteration in disease. Roman J Internal Med = Revue roumaine de medecine interne 35:3–11Google Scholar
  13. 13.
    Callewaert N, Van Vlierberghe H, Van Hecke A, Laroy W, Delanghe J, Contreras R (2004) Noninvasive diagnosis of liver cirrhosis using DNA sequencer-based total serum protein glycomics. Nat Med 10:429–434PubMedCrossRefGoogle Scholar
  14. 14.
    Liu XE, Desmyter L, Gao CF, Laroy W, Dewaele S, Vanhooren V, Wang L, Zhuang H, Callewaert N, Libert C, Contreras R, Chen C (2007) N-glycomic changes in hepatocellular carcinoma patients with liver cirrhosis induced by hepatitis b virus. Hepatology (Baltimore, Md) 46:1426–1435CrossRefGoogle Scholar
  15. 15.
    Freeze HH, Aebi M (2005) Altered glycan structures: The molecular basis of congenital disorders of glycosylation. Curr Opin Struct Biol 15:490–498PubMedCrossRefGoogle Scholar
  16. 16.
    Kobata A (2003) Glycobiology in the field of aging research–introduction to glycogerontology. Biochimie 85:13–24PubMedCrossRefGoogle Scholar
  17. 17.
    Vanhooren V, Desmyter L, Liu XE, Cardelli M, Franceschi C, Federico A, Libert C, Laroy W, Dewaele S, Contreras R, Chen C (2007) N-glycomic changes in serum proteins during human aging. Rejuvenation Res 10:521–531aPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Valerie Vanhooren
    • 1
    • 2
  • Wouter Laroy
    • 1
    • 2
  • Claude Libert
    • 1
    • 2
  • Cuiying Chen
    • 1
    • 2
    Email author
  1. 1.Department for Molecular Biomedical ResearchVIBGhentBelgium
  2. 2.Department of Molecular BiologyGhent UniversityGhentBelgium

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