The degradation of glycoproteins with lithium borohydride: Isolation and analysis ofO-glycopeptides with reducedC-terminal amino acid residue

Autor: Brusov Os, N. K. Kochetkov, Arbatskiĭ Np, Leonid M. Likhosherstov, Serebriakova Mv, Derevitskaia Va, Vladimir N. Shibaev
Rok vydání: 2000
Předmět:
Zdroj: Russian Journal of Bioorganic Chemistry. 26:45-53
ISSN: 1573-9163
1068-1620
DOI: 10.1007/bf02758860
Popis: By the example of fetuin and a blood-group-specific mucin from porcine stomach, we showed that, under conditions of reductive degradation of glycoproteins with LiBH4-LiOH in 70% aqueous tert-butyl alcohol, the reduction and cleavage of amide bonds occur much faster than the simultaneous beta-elimination of carbohydrate chains O-linked with Ser and Thr residues of the peptide chain. The major degradation products containing the O-linked glycans are the O-glycosylated derivatives of 2-aminopropane-1,3-diol and 2-aminobutane-1,3-diol (the products of reduction of glycosylated Ser and Thr) and the glycopeptides containing 2-4 amino acid residues with reduced C-terminal amino acid. Seventeen homogeneous O-glycopeptides were isolated from the fetuin degradation products by ion-exchange and reversed-phase HPLC. Their structures were determined by MALDI-TOF mass spectrometry and by analyses for amino acids, amino alcohols, and carbohydrates. The application of the reaction for characterization of O-glycans and localization of O-glycosylation sites in O- and N,O-glycoproteins is discussed.
Databáze: OpenAIRE
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