Document Detail


Characterization of oligosaccharide moieties of intact glycoproteins by microwave-assisted partial acid hydrolysis and mass spectrometry.
MedLine Citation:
PMID:  16124037     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A method, which utilizes microwave-assisted partial acid hydrolysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS), to elucidate oligosaccharide composition of intact glycoproteins is presented here. Glycoproteins, such as ribonuclease B, avidin, alpha1-acid glycoprotein, and fetuin, are used as model systems to demonstrate this technique. Partial cleavage of oligosaccharides from whole intact glycoproteins with trifluoroacetic acid was observed after a short exposure to microwaves. Due to the high-resolution mass spectra obtained by MALDI-TOFMS from glycoproteins with molecular weights less than 20 kDa, the compositions of oligosaccharides are readily derived for ribonuclease B and avidin. The data agree with the proposed oligosaccharide structures of ribonuclease B (five glycoforms) and avidin (eight glycoforms). Larger glycoproteins such as alpha1-acid glycoprotein (many glycoforms) and fetuin (many glycoforms) exhibited only broad peaks with no glycoform resolution. Nevertheless, this method can be used successfully for analysis of glycoproteins with molecular weights greater than 20 kDa to determine the presence or absence of glycosylation.
Authors:
Bao-Shiang Lee; Sangeeth Krishnanchettiar; Syed Salman Lateef; Nabila Salman Lateef; Shalini Gupta
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Rapid communications in mass spectrometry : RCM     Volume:  19     ISSN:  0951-4198     ISO Abbreviation:  Rapid Commun. Mass Spectrom.     Publication Date:  2005  
Date Detail:
Created Date:  2005-09-05     Completed Date:  2006-03-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8802365     Medline TA:  Rapid Commun Mass Spectrom     Country:  England    
Other Details:
Languages:  eng     Pagination:  2629-35     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2005 John Wiley & Sons, Ltd.
Affiliation:
Protein Research Laboratory, Research Resources Center, 835 S. Wolcott Avenue, University of Illinois, Chicago, IL 60612, USA. boblee@uic.edu
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MeSH Terms
Descriptor/Qualifier:
Acids / chemistry
Amino Acid Sequence
Animals
Avidin / chemistry
Egg White
Glycoproteins / chemistry*
Hydrogen-Ion Concentration
Hydrolysis
Microwaves*
Molecular Sequence Data
Oligosaccharides / analysis*,  chemistry*
Ribonucleases / chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Chemical
Reg. No./Substance:
0/Acids; 0/Glycoproteins; 0/Oligosaccharides; 1405-69-2/Avidin; EC 3.1.-/Ribonucleases; EC 3.1.27.-/ribonuclease B

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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