Skip to main content
SpringerLink
Log in

N-Glycan profiling in the study of human aging

  • Methods
  • Published:
Biogerontology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

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

References

  1. Varki A (1999) Essentials of glycobiology. Cold spring Harbor Laboratory Press

  2. Rudd PM, Dwek RA (1997) Rapid, sensitive sequencing of oligosaccharides from glycoproteins. Curr Opin Biotechnol 8:488–497

    Article  PubMed  CAS  Google Scholar 

  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 240:210–226

    Article  PubMed  CAS  Google Scholar 

  4. Suzuki S, Honda S (1998) A tabulated review of capillary electrophoresis of carbohydrates. Electrophoresis 19:2539–2560

    Article  PubMed  CAS  Google Scholar 

  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 250:82–101

    Article  PubMed  CAS  Google Scholar 

  6. Callewaert N, Geysens S, Molemans F, Contreras R (2001) Ultrasensitive profiling and sequencing of n-linked oligosaccharides using standard DNA-sequencing equipment. Glycobiology 11:275–281

    Article  PubMed  CAS  Google Scholar 

  7. Laroy W, Contreras R, Callewaert N (2006) Glycome mapping on DNA sequencing equipment. Nat Protoc 1:397–405

    Article  PubMed  CAS  Google Scholar 

  8. Kukuruzinska MA, Lennon K (1998) Protein n-glycosylation: molecular genetics and functional significance. Crit Rev Oral Biol Med 9:415–448

    Article  PubMed  CAS  Google Scholar 

  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 2:817–824

    Article  PubMed  CAS  Google Scholar 

  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):261–275

    PubMed  CAS  Google Scholar 

  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 28:1531–1536

    PubMed  CAS  Google Scholar 

  12. Matei L (1997) Plasma proteins glycosylation and its alteration in disease. Roman J Internal Med = Revue roumaine de medecine interne 35:3–11

    CAS  Google Scholar 

  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 10:429–434

    Article  PubMed  CAS  Google Scholar 

  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) 46:1426–1435

    Article  CAS  Google Scholar 

  15. Freeze HH, Aebi M (2005) Altered glycan structures: The molecular basis of congenital disorders of glycosylation. Curr Opin Struct Biol 15:490–498

    Article  PubMed  CAS  Google Scholar 

  16. Kobata A (2003) Glycobiology in the field of aging research–introduction to glycogerontology. Biochimie 85:13–24

    Article  PubMed  CAS  Google Scholar 

  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 10:521–531a

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr. Amin Bredan for editing the manuscript.

Author information

Authors and Affiliations

  1. Department for Molecular Biomedical Research, VIB, Technologiepark 927, 9052, Ghent, Belgium

    Valerie Vanhooren, Wouter Laroy, Claude Libert & Cuiying Chen

  2. Department of Molecular Biology, Ghent University, Technologiepark 927, 9052, Ghent, Belgium

    Valerie Vanhooren, Wouter Laroy, Claude Libert & Cuiying Chen

Authors
  1. Valerie Vanhooren
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wouter Laroy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Claude Libert
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cuiying Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cuiying Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vanhooren, V., Laroy, W., Libert, C. et al. N-Glycan profiling in the study of human aging. Biogerontology 9, 351–356 (2008). https://doi.org/10.1007/s10522-008-9140-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10522-008-9140-z

Keywords

Search

Navigation