期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1103, 期 2, 页码 296-306出版社
ELSEVIER
DOI: 10.1016/j.chroma.2005.11.043
关键词
mass spectrometric oligosaccharide profiling; graphitized carbon column; ion trap mass spectrometry; Fourier transform ion cyclotron resonance mass spectrometry; data-dependent MSn; Thy-1
We have previously described the site-specific glycosylation analysis of rat brain Thy-1 by LC/multistage tandem mass spectrometry (MS') using proteinase-digested Thy-1. In the present study, detailed structures of oligosaccharides released from Thy-1 were elucidated by mass spectrometric oligosaccharide profiling using LC/MS with a graphitized carbon column (GCC-LC/MS). First, using model oligosaccharides, we improved the oligosaccharide profiling by ion trap mass spectrometry (IT-NIS) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Sequential scanning of a full MSn scan with FT-ICR-MS followed by data-dependent MS' with IT-MS in positive ion mode, and a subsequent full MS1 scan with FT-ICR-MS followed by data-dependent MSn with IT-MS in negative ion mode enabled the monosaccharide composition analysis as well as profiling and sequencing of both neutral and acidic oligosaccharides in a single analysis. The improved oligosaccharide profiling was applied to elucidation of N-linked oligosaccharides from Thy-1 isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was demonstrated that Thy-1 possesses a significant variety of N-linked oligosaccharides, including Lewis a/x, Lewis b/y, and disialylated structure as a partial structure. Our method could be applicable to analysis of a small abundance of glycoproteins, and could become a powerful tool for glycoproteomics. (c) 2005 Elsevier B.V. All rights reserved.
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