Derivatization using dimethylamine for tandem mass spectrometric structure analysis of enzymatically and acidically depolymerized methyl cellulose
2005 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 77, no 9, 2948-2959 p.Article in journal (Refereed) Published
Structure analysis of partially depolymerized methyl cellulose was performed by nanoelectrospray ionization tandem mass spectrometry (nano-ESI-MS/MS) and by matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-MS/MS). Dimethylamine (DMA) was used for the first time as a reducing end derivatization reagent for oligosaccharides. This is an attractive reagent since it could be easily removed from the reaction mixture. Most important it also introduces a basic functional group that increased the sensitivity in both MALDI and nano-ESI. Depolymerization was made in two ways: one by the cellulose selective endoglucanase 5A from Bacillus agaradhaerens (Ba Ce15A) and the other by trifluoroacetic acid. The DMA derivatives formed both protonated and sodiated molecules in nano-ESI and MALDI. Tandem MS of protonated molecules yielded predominantly Y fragments from which the distribution of the substituents in the oligomers could be measured. Fragments obtained in tandem MS of sodiated molecules provided information regarding the positions of the substituents within the anhydroglucose units (AGUs). It was found that Ba Ce15A could cleave glucosidic bonds also if the AGU on the reducing side of the bond was fully methylated. The combination of DMA derivatization and tandem MS was demonstrated as a tool for the characterization of endoglucanase selectivity.
Place, publisher, year, edition, pages
2005. Vol. 77, no 9, 2948-2959 p.
collision-induced dissociation, electrospray-ionization, substituent distribution, carboxymethyl cellulose, glycosyl hydrolases, reaction coordinate, residue loss, fab-ms, oligosaccharides, sequence
IdentifiersURN: urn:nbn:se:kth:diva-14725DOI: 10.1021/ac048194eISI: 000228877500047OAI: oai:DiVA.org:kth-14725DiVA: diva2:332766
QC 201005252010-08-052010-08-052014-04-02Bibliographically approved