Summary
Human cerebrospinal fluid (CSF), which circulates within the ventricles of the brain and the subarachnoid space of the central nervous system (CNS), is an excellent source for proteomic discovery of biomarkers in neurodegenerative disorders, including Alzheimer’s and Parkinson’s disease. Protein glycosylation is an abundant and biologically significant posttranslational modification. Glycoproteins, commonly associated with membrane and secreted proteins, are highly enriched in body fluids, including CSF. Focusing on glycoproteins also improves the dynamic range of proteomic profiling of the CSF, where low abundance proteins are difficult to identify because of the CSF’s enormous complexity. As an ongoing process to define the human CSF proteome, we have recently employed a complementary proteomic approach, with integrated lectin affinity column and hydrazide chemistry, for CSF glycoprotein identification. This investigation has revealed many proteins of low abundance that are related to the CNS structurally and/or functionally. This review centers on the technical details involved in various steps in sample preparation as well as proteomic analysis of CSF glycoproteins.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Blennow, K. (2005) CSF biomarkers for Alzheimer’s disease: Use in early diagnosis and evaluation of drug treatment. Expert. Rev. Mol. Diagn. 5, 661–672.
Zhang, J., Goodlett, D. R., and Montine, T. J. (2005) Proteomic biomarker discovery in cerebrospinal fluid for neurodegenerative diseases. J. Alzheimers Dis. 8, 377–386.
Gronborg, M., Kristiansen, T. Z., Stensballe, A., Andersen, J. S., Ohara, O., Mann, M., Jensen, O. N., and Pandey, A. (2002) A mass spectrometry-based proteomic approach for identification of serine/threonine-phosphorylated proteins by enrichment with phospho-specific antibodies: Identification of a novel protein, Frigg, as a protein kinase A substrate. Mol. Cell Proteomics 1, 517–527.
Yang, Z., and Hancock W. S. (2004) Approach to the comprehensive analysis of glycoproteins isolated from human serum using a multi-lectin affinity column. J. Chromatogr. A 1053, 79–88.
Liu, T., Qian, W. J., Gritsenko, M. A., Camp, D. G., 2nd, Monroe, M. E., Moore, R. J., and Smith, R. D. (2005) Human plasma N-glycoproteome analysis by immunoaffinity subtraction, hydrazide chemistry, and mass spectrometry. J. Proteome. Res. 4, 2070–2080.
Yang, Z., Hancock, W. S., Chew, T. R., and Bonilla, L. (2005) A study of glycoproteins in human serum and plasma reference standards (HUPO) using multilectin affinity chromatography coupled with RPLC-MS/MS. Proteomics 5, 3353–3366.
Jaeken, J., and Matthijs, G. (2001) Congenital disorders of glycosylation. Annu. Rev. Genomics Hum. Genet. 2, 129–151.
Rudd, P. M., Elliott, T., Cresswell, P., Wilson, I. A., and Dwek, R. A. (2001) Glycosylation and the immune system. Science 291, 2370–2376.
Huang, X., Ushijima, K., Komai, K., Takemoto, Y., Motoshima, S., Kamura, T., and Kohno, K. (2004) Co-expression of Y box-binding protein-1 and P-glycoprotein as a prognostic marker for survival in epithelial ovarian cancer. Gynecol. Oncol. 93, 287–291.
Ferrara, N., and Kerbel, R. S. (2005) Angiogenesis as a therapeutic target. Nature 438, 967–974.
Burton, D. R., and Dwek, R. A. (2006) Immunology. Sugar determines antibody activity. Science 313, 627–628.
Geng, M., Zhang, X., Bina, M., and Regnier, F. (2001) Proteomics of glycoproteins based on affinity selection of glycopeptides from tryptic digests. J. Chromatogr. B Biomed. Sci. Appl. 752, 293–306.
Kaji, H., Saito, H., Yamauchi, Y., Shinkawa, T., Taoka, M., Hirabayashi, J., Kasai, K., Takahashi, N., and Isobe, T. (2003) Lectin affinity capture, isotope-coded tagging and mass spectrometry to identify N-linked glycoproteins. Nat. Biotechnol. 21, 667–672.
Zhang, H., Li, X. J., Martin, D. B., and Aebersold, R. (2003) Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry. Nat. Biotechnol. 21, 660–666.
Pan, S., Zhu, D., Quinn, J. F., Peskind, E. R., Montine, T. J., Lin, B., et al. (2007) A combined dataset of human cerebrospinal fluid proteins identified by multi-dimensional chromatography and tandem mass spectrometry. Proteomics 7, 469–473.
Pan, S., Wang, Y., Quinn, J. F., Peskind, E. R., Waichunas, D., Wimberger, J. T., Jin, J., Li, J. G., Zhu, D., Pan, C., and Zhang, J. (2006) Identification of glycoproteins in human cerebrospinal fluid with a complementary proteomic approach. J. Proteome. Res. 5, 2769–2779.
Gatlin, C. L., Eng, J. K., Cross, S. T., Detter, J. C., and Yates, J. R. 3rd. (2000) Automated identification of amino acid sequence variations in proteins by HPLC/microspray tandem mass spectrometry. Anal. Chem. 72, 757–763.
Keller, A., Nesvizhskii, A. I., Kolker, E., and Aebersold, R. (2002) Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal. Chem. 74, 5383–5392.
Nesvizhskii, A. I., Keller, A., Kolker, E., and Aebersold, R. (2003) A statistical model for identifying proteins by tandem mass spectrometry. Anal. Chem. 75, 4646–4658.
Zhang, J. (2007) Proteomics of human cerebrospinal fluid – the good, the bad, and the ugly. Proteomics Clin. Appl. 1, 805–819.
Blennow, K., Fredman, P., Wallin, A., Gottfries, C. G., Langstrom, G., and Svennerholm, L. (1993) Protein analyses in cerebrospinal fluid. I. Influence of concentration gradients for proteins on cerebrospinal fluid/serum albumin ratio. Eur. Neurol. 33, 126–128.
Blennow, K., Fredman, P., Wallin, A., Gottfries, C. G., Skoog, I., Wikkelso, C., et al. (1993) Protein analysis in cerebrospinal fluid. III. Relation to blood–cerebrospinal fluid barrier function for formulas for quantitative determination of intrathecal IgG production. Eur. Neurol. 33, 134–142.
Blennow, K., Fredman, P., Wallin, A., Gottfries, C. G., Karlsson, I., Langstrom, G., Skoog, I., Svennerholm, L., and Wikkelsö, C. (1993) Protein analysis in cerebrospinal fluid. II. Reference values derived from healthy individuals 18–88 years of age. Eur. Neurol. 33, 129–133.
Ding, W., Hill, J. J., and Kelly, J. (2007) Selective enrichment of glycopeptides from glycoprotein digests using ion-pairing normal-phase liquid chromatography. Anal. Chem. 79, 8891–8899.
Acknowledgments
The research is supported by following grants from the NIH (AG025327, ES012703, and NS060252) as well as the Michael J. Fox Foundation and a Shaw Endowment to Dr. Jing Zhang.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Hwang, H.J., Quinn, T., Zhang, J. (2009). Identification of Glycoproteins in Human Cerebrospinal Fluid. In: Ottens, A., Wang, K. (eds) Neuroproteomics. Methods in Molecular Biology, vol 566. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-562-6_17
Download citation
DOI: https://doi.org/10.1007/978-1-59745-562-6_17
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-934115-84-8
Online ISBN: 978-1-59745-562-6
eBook Packages: Springer Protocols