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Discrimination of GalNAc (4S/6S) sulfation sites in chondroitin sulfate disaccharides by chip-based nanoelectrospray multistage mass spectrometry

Domenii publicaţii > Chimie + Tipuri publicaţii > Articol în revistã ştiinţificã

Autori: C.Flangea, A.F.Serb, C.Schiopu1, S.Tudor, E.Sisu, D.G.Seidler, A.D.Zamfir

Editorial: Springer, Cent. Eur. J. Chem., 7(4), p.752-759, 2009.


Sulfation pattern within chondroitin sulfate (CS) glycosaminoglycan (GAG) chains is an important post-translational modification that
regulates their interaction with proteins. In this context, development of highly efficient and reproducible analytical methods for the
investigation of CS sulfation patterns is of high necessity. In this study we report a novel method for straightforward determination of
N-acetylgalactosamine (GalNAc) sulfation sites in chondroitin sulfate disaccharides. Our protocol involves combining fully automated
chip-based nanoelectrospray (nanoESI) for analyte infusion and ionization in negative ion mode with multistage (MSn) collision-induced
dissociation (CID) high capacity ion trap (HCT) mass spectrometry for generation of sequence ions diagnostic for identification of
sulfate ester group position within GalNAc residues. The feasibility of this approach is here demonstrated on chondroitin 6-O-sulfate
and chondroitin 4-O-sulfate disaccharides. Fragmentation patterns obtained by MS2 and MS3 sequencing stages provided first mass
spectrometric data from which sulfation site(s) within GalNAc monosaccharide ring could be unequivocally deciphered. Hence, the
method allowed discriminating 4S/6S sulfation sites solely on the basis of MS and multistage MS evidence.

Cuvinte cheie: Condroitinsulfat.Profil de sulfatare.Infuzie automata prin chip nano-ESI..Spectrometrie de masa de inalta capacitate cu trapa ionica // Chondroitin sulfate