Characterization of protein glycosylation by mass spectrometry.
Electrospray ionization, a natural interface with microbore and capillary high-pressure liquid chromatography, has become the method of choice for the reliable structural characterization of protein glycosylation by mass spectrometry at the picomole level. Its advantages include inherent sensitivity in the femtomole range, compatibility with collisional activation methods that both permit the detection and monitoring of structurally specific ions and enable the induction of glycopeptide fragmentation that facilitates determination of glycoform sequence and branching. Developments in high-performance electrospray mass spectrometry include sample introduction at nanoliter flow rates, tandem magnetic sector/orthogonal time-of-flight instruments, Fourier transform instruments, and new ion optical strategies, including ion traps. Although a sensitive and important complementary technique, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry suffers from matrix-dependent deposition of excess internal energies, which produce extensive metastable fragmentation and (photo)adduct formation. These metastable fragments may be focused into a mass spectrum by employing an ion mirror (reflectron) in time-of-flight instrumentation. In favorable cases, structural information may be obtained.
Burlingame
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Department of Pharmaceutical Chemistry and the Liver Center, University of California, San Francisco 94143-0446, USA.
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005583:Fourier Analysis; D006020:Glycopeptides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008563:Membrane Lipids / Q000032:analysis; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D012680:Sensitivity and Specificity
D006020:Glycopeptides; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D008563:Membrane Lipids; D009844:Oligosaccharides; D011506:Proteins