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Gewählte Publikation:

Figl, R; Altmann, F.
(2018): Reductive Alkaline Release of N-Glycans Generates a Variety of Unexpected, Useful Products
PROTEOMICS. 2018; 18(3-4): FullText FullText_BOKU

Abstract:
Release of O-glycans by reductive -elimination has become routine in many glyco-analytical laboratories and concomitant release of N-glycans has repeatedly been observed. Revisiting this somewhat forgotten mode of N-glycan release revealed that all kinds of N-glycans including oligomannosidic and complex-type N-glycans from plants with 3-linked fucose and from mammals with or without 6-linked fucose and with sialic acid could be recovered. However, the mass spectra of the obtained products revealed very surprising facts. Even after 16h incubation in 1M sodium borohydride, a large part of the glycans occurred in reducing form. Moreover, about one third emerged in the form of the stable amino-functionalized 1-amino-1-deoxy-glycitol. When avoiding acidic conditions, considerable amounts of glycosylamine were observed. In addition, a compound with a reduced asparagine and de-N-acetylation products, in particular of sialylated glycans, was seen. The relative yields of the products reducing glycosylamine, reducing N-glycan, 1-amino-1-deoxy-glycitol or glycitol could be controlled by the release conditions, foremost by temperature and borohydride concentration. Thus, chemical release of N-glycans constitutes a cost-saving alternative to enzymatic hydrolysis for the preparation of precursors for the production of reference compounds for various formats of N-glycan analysis. Moreover, it allows to obtain a stable amino-functionalized glycan derivative, which can be employed to construct glycan arrays or affinity matrices.
Autor/innen der BOKU Wien:
Altmann Friedrich
Figl Rudolf
BOKU Gendermonitor:


Find related publications in this database (Keywords)
glycoprotein
isotope pattern
mass spectrometry
N-glycan
reductive alkaline release


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