One‐pot nonreductive O‐glycan release and labeling with 1‐phenyl‐3‐methyl‐5‐pyrazolone followed by ESI‐MS analysis
Identifieur interne : 000D26 ( Main/Exploration ); précédent : 000D25; suivant : 000D27One‐pot nonreductive O‐glycan release and labeling with 1‐phenyl‐3‐methyl‐5‐pyrazolone followed by ESI‐MS analysis
Auteurs : Chengjian Wang [République populaire de Chine] ; Wancui Fan [République populaire de Chine] ; Ping Zhang [République populaire de Chine] ; Zhongfu Wang [République populaire de Chine] ; Linjuan Huang [République populaire de Chine]Source :
- PROTEOMICS [ 1615-9853 ] ; 2011-11.
English descriptors
- Teeft :
- Alditols, Ammonia, Ammonia solution, Anal, Aqueous ammonia, Aqueous methanol, Biochem, Borohydride, Bovine, Bovine fetuin, Capillary electrophoresis, Carbohydrate, Chem, Derivative, Derivatization, Electrospray, Eluate, Fetuin, Glycan, Glycans, Glycoprotein, Gmbh, Good stability, Graphitized, Graphitized carbon, Hexosamine, High sensitivity, Hplc, Huang, Human plasma, Kgaa, Liquid chromatography, Maltohexaose, Mass spectrom, Mass spectrometry, Mild conditions, Molecular weights, Monosaccharide, Mucin, Mucin glycans, Nonreductive, Nonreductive release, Oligosaccharide, Original glycans, Peak intensity, Peak intensity ratios, Peptide, Porcine, Porcine stomach mucin, Porcine stomach mucin type, Proteomics, Reaction conditions, Reaction temperature, Reaction time, Reductive, Ribonuclease, Saccharide, Sodium borohydride, Spectrometry, Structural analysis, Structure procedure, Traditional reductive, Verlag, Verlag gmbh, Wang, Weinheim, Weinheim proteomics, Whole blood plasma, Zhang.
Abstract
A novel one‐pot procedure for the nonreductive release of O‐linked glycans from glycoproteins and the simultaneous derivatization of released glycans with 1‐phenyl‐3‐methyl‐5‐pyrazolone (PMP) is described. Unlike the traditional reductive β‐elimination, which produces alditols, this new method employs PMP/ammonia aqueous solution as the reaction medium. The O‐glycans are released from glycoproteins and derivatized with PMP nonreductively, specifically, and quantitatively. Samples can be easily purified from ammonia, excess PMP, and peptide residues by evaporation, chloroform extraction, and solid‐phase extraction (SPE) column fractionation for HPLC, CE, or MS analysis. The procedure has been elaborated with two purified glycoproteins, porcine stomach mucin and bovine fetuin, and successfully applied to O‐glycan profiling of a challenging biological specimen, healthy human plasma. This new procedure has shown methodological significance in O‐glycan analysis.
Url:
DOI: 10.1002/pmic.201000677
Affiliations:
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plasma</term><term>Kgaa</term><term>Liquid chromatography</term><term>Maltohexaose</term><term>Mass spectrom</term><term>Mass spectrometry</term><term>Mild conditions</term><term>Molecular weights</term><term>Monosaccharide</term><term>Mucin</term><term>Mucin glycans</term><term>Nonreductive</term><term>Nonreductive release</term><term>Oligosaccharide</term><term>Original glycans</term><term>Peak intensity</term><term>Peak intensity ratios</term><term>Peptide</term><term>Porcine</term><term>Porcine stomach mucin</term><term>Porcine stomach mucin type</term><term>Proteomics</term><term>Reaction conditions</term><term>Reaction temperature</term><term>Reaction time</term><term>Reductive</term><term>Ribonuclease</term><term>Saccharide</term><term>Sodium borohydride</term><term>Spectrometry</term><term>Structural analysis</term><term>Structure procedure</term><term>Traditional reductive</term><term>Verlag</term><term>Verlag gmbh</term><term>Wang</term><term>Weinheim</term><term>Weinheim proteomics</term><term>Whole blood plasma</term><term>Zhang</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel one‐pot procedure for the nonreductive release of O‐linked glycans from glycoproteins and the simultaneous derivatization of released glycans with 1‐phenyl‐3‐methyl‐5‐pyrazolone (PMP) is described. Unlike the traditional reductive β‐elimination, which produces alditols, this new method employs PMP/ammonia aqueous solution as the reaction medium. The O‐glycans are released from glycoproteins and derivatized with PMP nonreductively, specifically, and quantitatively. Samples can be easily purified from ammonia, excess PMP, and peptide residues by evaporation, chloroform extraction, and solid‐phase extraction (SPE) column fractionation for HPLC, CE, or MS analysis. The procedure has been elaborated with two purified glycoproteins, porcine stomach mucin and bovine fetuin, and successfully applied to O‐glycan profiling of a challenging biological specimen, healthy human plasma. This new procedure has shown methodological significance in O‐glycan analysis.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Chengjian" sort="Wang, Chengjian" uniqKey="Wang C" first="Chengjian" last="Wang">Chengjian Wang</name></noRegion><name sortKey="Fan, Wancui" sort="Fan, Wancui" uniqKey="Fan W" first="Wancui" last="Fan">Wancui Fan</name><name sortKey="Huang, Linjuan" sort="Huang, Linjuan" uniqKey="Huang L" first="Linjuan" last="Huang">Linjuan Huang</name><name sortKey="Huang, Linjuan" sort="Huang, Linjuan" uniqKey="Huang L" first="Linjuan" last="Huang">Linjuan Huang</name><name sortKey="Wang, Zhongfu" sort="Wang, Zhongfu" uniqKey="Wang Z" first="Zhongfu" last="Wang">Zhongfu Wang</name><name sortKey="Wang, Zhongfu" sort="Wang, Zhongfu" uniqKey="Wang Z" first="Zhongfu" last="Wang">Zhongfu Wang</name><name sortKey="Zhang, Ping" sort="Zhang, Ping" uniqKey="Zhang P" first="Ping" last="Zhang">Ping Zhang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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