LC/MSn for glycoprotein analysis: N-linked glycosylation analysis and peptide sequencing of glycopeptides.
Liquid chromatography/multiple-stage mass spectrometry (LC/MS( n )) is an effective means for the site-specific glycosylation analysis of a limited quantity of glycoproteins, such as gel-separated proteins. Generally, a tryptic digest of the glycoprotein is separated by reversed-phase LC, and peptide sequencing and glycosylation analysis are achieved with on-line MS( n ). In this chapter, a protocol for the LC/MS/MS/MS of a proteolytic digest of a gel-separated glycoprotein is described.
Kawasaki
Nana
N
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. nana@nihs.go.jp
Itoh
Satsuki
S
Yamaguchi
Teruhide
T
D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational / Q000502:physiology; D019163:Reducing Agents / Q000494:pharmacology; D020539:Sequence Analysis, Protein / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D006023:Glycoproteins; D019163:Reducing Agents