Titanium dioxide enrichment of sialic acid-containing glycopeptides

Giuseppe Palmisano, Sara Eun Lendal, Martin R. Larsen

Research output: Chapter in Book/Report/Conference proceedingChapter

22 Citations (Scopus)

Abstract

Glycosylation is one of the many post-translational protein modifications that regulate several biological processes of proteins and lipids. In particular aberrant sialylation, at the terminal position of the glycan structures of cell surface proteins, occurs in numerous diseases such as cancer metastasis and viral infections. Methodological improvements in the sample preparation and analysis currently enable the detailed identification of the glycosylation sites and glycan structure characterization. In this context, the aim of this chapter is to describe a methodology to identify the glycosylation site of N-linked sialylated glycoproteins. The method relies on the specificity of titanium dioxide affinity chromatography to isolate sialic acid-containing glycopeptides. After enzymatic release of the glycans, the enriched sialylated glycopeptides are analyzed by mass spectrometry. This strategy was applied to a crude membrane fraction of EGF-stimulated HeLa cells metabolically labeled with SILAC enabling both qualitative and quantitative analyses of sialoglycopeptides.

Original languageEnglish
Title of host publicationGel-Free Proteomics
Subtitle of host publicationMethods and Protocols
Pages309-322
Number of pages14
DOIs
Publication statusPublished - 1 Dec 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume753
ISSN (Print)1064-3745

Fingerprint

Glycopeptides
N-Acetylneuraminic Acid
Glycosylation
Polysaccharides
Sialoglycoproteins
Biological Phenomena
Virus Diseases
Post Translational Protein Processing
Affinity Chromatography
HeLa Cells
Epidermal Growth Factor
Mass Spectrometry
Glycoproteins
Membrane Proteins
Neoplasm Metastasis
Lipids
Membranes
titanium dioxide
Neoplasms
Proteins

Keywords

  • Glycosylation
  • Mass spectrometry
  • Quantitative proteomics
  • Sialylation
  • Titanium dioxide

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Palmisano, G., Lendal, S. E., & Larsen, M. R. (2011). Titanium dioxide enrichment of sialic acid-containing glycopeptides. In Gel-Free Proteomics: Methods and Protocols (pp. 309-322). (Methods in Molecular Biology; Vol. 753). https://doi.org/10.1007/978-1-61779-148-2_21

Titanium dioxide enrichment of sialic acid-containing glycopeptides. / Palmisano, Giuseppe; Lendal, Sara Eun; Larsen, Martin R.

Gel-Free Proteomics: Methods and Protocols. 2011. p. 309-322 (Methods in Molecular Biology; Vol. 753).

Research output: Chapter in Book/Report/Conference proceedingChapter

Palmisano, G, Lendal, SE & Larsen, MR 2011, Titanium dioxide enrichment of sialic acid-containing glycopeptides. in Gel-Free Proteomics: Methods and Protocols. Methods in Molecular Biology, vol. 753, pp. 309-322. https://doi.org/10.1007/978-1-61779-148-2_21
Palmisano G, Lendal SE, Larsen MR. Titanium dioxide enrichment of sialic acid-containing glycopeptides. In Gel-Free Proteomics: Methods and Protocols. 2011. p. 309-322. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-148-2_21
Palmisano, Giuseppe ; Lendal, Sara Eun ; Larsen, Martin R. / Titanium dioxide enrichment of sialic acid-containing glycopeptides. Gel-Free Proteomics: Methods and Protocols. 2011. pp. 309-322 (Methods in Molecular Biology).
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