Simultaneous glycosylation analysis of human serum glycoproteins by high-performance liquid chromatography/tandem mass spectrometry.
Changes in the glycosylation of some serum proteins are associated with certain diseases. In this study, we performed simultaneous site-specific glycosylation analysis of abundant serum glycoproteins by LC/Qq-TOF MS of human serum tryptic digest, the albumin of which was depleted. The glycopeptide peaks on the chromatogram were basically assigned by database searching with modified peak-list text files of MS/MS spectra and then based on mass differences of glycan units from characterized glycopeptides. Glycopeptide of IgG, haptoglobin and ceruloplasmin were confirmed by means of a comparison of their retention times and m/z values with those obtained by LC/MS of commercially available glycoproteins. Mass spectrometric carbohydrate heterogeneity in the assigned glycopeptides was analyzed by an additional LC/MS. We successfully demonstrated site-specific glycosylation of 23 sites in abundant serum glycoproteins.
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo, Japan. harazono@nihs.go.jp
Akira
A
Harazono
Nana
N
Kawasaki
Satsuki
S
Itoh
Noritaka
N
Hashii
Yukari
Y
Matsuishi-Nakajima
Toru
T
Kawanishi
Teruhide
T
Yamaguchi
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D030562:Databases, Protein; D006020:Glycopeptides / Q000032:analysis / Q000302:isolation & purification; D006023:Glycoproteins / Q000097:blood* / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D011499:Protein Processing, Post-Translational; D053719:Tandem Mass Spectrometry / Q000379:methods*
D006020:Glycopeptides; D006023:Glycoproteins