Structural characterization of chondroitin/dermatan sulfate oligosaccharides from bovine aorta by capillary electrophoresis and electrospray ionization quadrupole time-of-flight tandem mass spectrometry.
An analytical approach based on high-performance capillary electrophoresis (CE) in conjunction with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-QTOF-MS/MS) has been developed for providing the basis to obtain new insights into the domain structure of the glycosaminoglycan (GAG) moiety of proteoglycans. The feasibility and performance of the off-line CE/ESI-QTOF-MS approach in GAG oligosaccharide analysis were assessed by screening a chondroitin/dermatan sulfate (DS) oligosaccharide mixture obtained from bovine aorta by enzymatic depolymerization by chondroitin B lyase. The CS/DS mixture was analyzed by CE using 50 mM ammonium acetate, pH 12.0, dissolved in aqueous methanol (2:3; v/v), as a CE carrier. Structural identification of the GAG components was achieved using off-line CE/nanoESI-QTOF-MS and-MS/MS experiments. ESI-QTOF instrumental parameters were found to play an important role in the MS screening of the CE-separated GAG species. By optimizing the ESI conditions, oligosaccharides differing in chain length and degree of sulfation could be detected. The building block composition, the size of the carbohydrate chain, as well as structural features of the repeating HexA-GalNAc, HexA-GalNAc(S) units, have been determined using MS/MS by applying collision-induced dissociation at low energies. Cleavage of GAG chains by chondroitin B lyase occurs with formation of structural markers useful for identification of IdoA-containing domains.
Zamfir
Alina
A
Institute for Medical Physics and Biophysics, Biomedical Analysis Department, University of Münster, Robert-Koch-Str. 31, D-48149 Münster, Germany.
Seidler
Daniela G
DG
Kresse
Hans
H
Peter-Katalinić
Jasna
J
D000818:Animals; D001011:Aorta / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002417:Cattle; D002807:Chondroitin / Q000737:chemistry*; D003871:Dermatan Sulfate / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005240:Feasibility Studies; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D009844:Oligosaccharides; D003871:Dermatan Sulfate; D002807:Chondroitin