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Capillary electrophoresis-electrospray mass spectrometry for the characterization of high-mannose-type N-glycosylation and differential oxidation in glycoproteins by charge reversal and protease/glycosidase digestion


, : Capillary electrophoresis-electrospray mass spectrometry for the characterization of high-mannose-type N-glycosylation and differential oxidation in glycoproteins by charge reversal and protease/glycosidase digestion. Analytical Chemistry 73(24): 5875-5885

The characterization of high-mannose-type N-glycosylation by capillary electrophoresis-electrospray mass spectrometry (CE-ESI MS) was described. In addition to the use of a cationic noncovalent capillary coating, strong acidic buffer, and charge reversal to increase the glycoform resolving power, N-glycosidase F (PNGase F) combined with a basic protease and alpha-mannosidase combined with an acidic protease were used to analyze the high-mannose-type N-glycosylation in ribonuclease B (RNase B) and in a novel C-type lectin from the venom of Trimeresurus stejnegeri (TSL). The structures of oligosaccharide, glycosylation sites, and glycoform distributions were determined simultaneously, and the differential oxidation of Met residues in glycopeptides obtained from TSL protease digestion was also characterized successfully by CE-MS/MS. The results showed that the oligosaccharide attached to RNase B has a structure of GlcNAc2Man5 approximately 9, and that attached to TSL has a structure of GlcNAc2Min5 approximately 8. The glycoform distributions in these glycoproteins are quite different, with the GlcNAc2Man5 type predominant in RNase B, and the GlcNAc2Man8 type, in TSL This method may be useful not only for the characterization of glycosylation sites and glycan structures, but also for the determination of the relative abundance of individual glycoforms.

US$29.90

PMID: 11791556


Other references

Boxho, P., 1997: The practice of placing under psychiatric observation. Revue Medicale de Liege 52(5): 345-351

Hare; Gross; Goy, 1988: Circular atoms prepared by a new method of crossed electric and magnetic fields. Physical Review Letters 61(17): 1938-1941

Cáliz, R.; del Amo, J.; Balsa, A.; Blanco, F.; Silva, L.; Sanmarti, R.; Martínez, F.G.; Collado, M.D.; Ramirez, M.del.Carmen.; Tejedor, D.; Artieda, M.; Pascual-Salcedo, D.; Oreiro, N.; Andreu, J.L.; Graell, E.; Simon, L.; Martínez, A.; Mulero, J., 2012: The C677T polymorphism in the MTHFR gene is associated with the toxicity of methotrexate in a Spanish rheumatoid arthritis population. Methotrexate (MTX) is the first-choice drug for the treatment of rheumatoid arthritis (RA) patients. However, 30% of RA patients discontinue therapy within 1 year, usually because of adverse effects. Previous studies have reported conflicting resu...

Boehlau, V., 1959: Influenza and physical efficiency. Verhandlungen der Deutschen Gesellschaft für Innere Medizin 65: 792-793

Lin, G-Ru.; Meng, F-Shuen.; Pai, Y-Hao.; Chang, Y-Chung.; Hsu, S-Hsin., 2010: Manipulative depolarization and reflectance spectra of morphologically controlled nano-pillars and nano-rods. Depolarization of sub-mum-high Si nano-pillar/nano-rod surface reflectance with morphologically controlled anti-reflection spectrum is demonstrated. Extremely small reflectance dip of 1.5% at 400-450 nm for Si nano-pillars is extraordinary when co...

Mehigan, D.G.; Bell, W.R.; Zuidema, G.D.; Eggleston, J.C.; Cameron, J.L., 1980: Disseminated intravascular coagulation and portal hypertension following pancreatic islet autotransplantation. A patient undergoing subtotal pancreatectomy and intraportal islet tissue autotransplantation for chronic pancreatitis developed severe portal hypertension (49 cm of H(2)O) and acute disseminated intravascular coagulation (DIC). In an attempt to i...

Ruiz-Moreno, J.M.; Montero, J.A.; Gomez-Ulla, F., 2009: Photodynamic therapy may worsen the prognosis of highly myopic choroidal neovascularisation treated by intravitreal bevacizumab. British Journal of Ophthalmology 93(12): 1693-1694

Farhat, K.; Riekenberg, S.; Jung, Günther.; Wiesmüller, K-Heinz.; Jungi, T.W.; Ulmer, A.J., 2010: Identification of full length bovine TLR1 and functional characterization of lipopeptide recognition by bovine TLR2/1 heterodimer. Toll-like receptors (TLR) are highly conserved pattern recognition receptors of the innate immune system. Toll-like receptor 2 (TLR2) recognizes bacterial lipopeptides in a heterodimeric complex with TLR6 or TLR1, thereby discriminating between di...

Sacktor, B.; Dick, A., 1962: Pathways of hydrogen transport in the oxidation of extramitochondrial reduced diphosphopyridine nucleotide in flight muscle. Teased flight muscle preparations of the blowfly, Phormla regina, do not oxidize added reduced diphosphopyridine nucleotide (DPNH) directly. This indicates a complete cytoplasmic-mitochondrial barrier to extramitochondrial DPNH. Alternative pathwa...

Graczykowska-Kocozowswka, A.; Tycowa, M.; Odrakiewicz, M.; Sokolnicka, B., 1970: Diet treatment of obesity. Out-patient treatment of obesity in 505 patients from 18 to 60 years old of whom 91% were women included a diet providing from 700 to 1400 kcal daily with protein 1.5 g/kg bodyweight in the form of egg, fat-free milk, lean meat, lean bacon and fis...