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. 2014 Dec 6;14:101. doi: 10.1186/s12896-014-0101-0

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© West et al.; licensee BioMed Central. 2014

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Glycoconjugate analysis of hGGT1 expressed in Pichia pastoris . (A) MALDI-TOF-MS profile of neutral N-glycans released by PNGase F from purified Pp-hGGT1. Ions corresponding to monosodiated N-glycans known to be expressed by Pichia pastoris are assigned. Symbols: blue box, GlcNAc; green circle, mannose. (B) Averaged LC/Q-Star MS spectrum for the Asn-120 (LAFATMFNSSEQSQK) family of glycopeptides from Pp-hGGT1, showing the identified glycoconjugates within their elution interval. Asterisks denote glycoconjugates that were confirmed by tandem MS analysis. P represents the LAFATMFNSSEQSQK tryptic peptide to which the N-glycans are attached. Unannotated peaks represent co-eluting nonglycosylated hGGT1 peptides. (C) MS/MS spectrum showing fragmentation of the Man₁₁GlcNAc₂-modified glycopeptide (m/z =1293.2) shown in (B).