Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Organic Chemistry Regular Papers
Mass Spectrometric Assignment of Smith Degradation Glycopeptides Derived from Ribonuclease B
Suthasinee PITCHAYAWASINMinoru ISOBE
Author information
JOURNAL FREE ACCESS

2004 Volume 68 Issue 7 Pages 1424-1433

Details
Abstract

We established a method to determine the glycosyl linkage structure by a combination of Smith degradation and liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-Q-TOF-MS) and tandem MS (MS/MS). To assign the sugar linkage of N-glycoprotein, we employed a typical ribonuclease B containing oligosaccharides (Man5–9GlcNAc2). Tryptic digestion of ribonuclease B provided a mixture of high-mannose glycopeptides consisting of the four amino acids, Asn34-Leu-Thr-Lys37 (NLTK, T6). The mixture of glycopeptides was separated by high-performance liquid chromatography (HPLC) in a reversed phase column and was characterized by ESI-Q-TOF-MS and MS/MS. Comparison of the data with and without Smith degradation allowed us to make reasonable assignments to support such linkage patterns as (1→2), (1→3), (1→6) and their multiples. These assignments were limited to six mannoses or lower due to the unstable nature of the higher derivatives. This method should be applicable to determine the linkage pattern of an unknown glycoprotein in about a 6-microgram amount.

Content from these authors

This article cannot obtain the latest cited-by information.

© 2004 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
Previous article Next article
feedback
Top