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Glycomic mapping of O- and N-linked glycans from major rat sublingual mucin

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Abstract

Carbohydrate moieties of salivary mucins play various roles in life processes, especially as a microbial trapping agent. While structural details of the salivary O-glycans from several mammalian sources are well studied, very little information is currently available for the corresponding N-glycans. The existence of N-glycans alongside O-glycans on mucin isolated from rat sublingual gland has previously been implicated by total glycosyl compositional analysis but the respective structural data are both lacking. The advent of facile glycomic mapping and sequencing methods by mass spectrometry (MS) has enabled a structural reinvestigation into many previously unsolved issues. For the first time, high energy collision induced dissociation (CID) MALDI-MS/MS as implemented on a TOF/TOF instrument was applied to permethyl derivatives of mucin type O-glycans and N-glycans, from which the linkage specific fragmentation pattern could be established. The predominant O-glycans carried on the rat sublingual mucin were defined as sialylated core 3 and 4 types whereas the N-glycans were determined to be non-bisected hybrid types similarly carrying a sialylated type II chain. The masking effect of terminal sialylation on the tight binding of rat sublingual mucin to Galβ1→4GlcNAc specific lectins and three oligomannose specific lectins were clearly demonstrated in this study.

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Abbreviations

ASG:

armadillo salivary gland

BSM:

bovine submandibular glycoprotein

ELLSA:

enzyme-linked lectinosorbent assay

OSM:

ovine submandibular glycoprotein

PSM:

porcine salivary glycoprotein

RSL:

rat sublingual glycoprotein

Tn:

GalNAcα1→Ser/Thr

type I:

Galβ1→3GlcNAc, or lacto-N-biose I

type II:

Galβ1→4GlcNAc, or N-acetyllactosamine

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Acknowledgement

This study was supported by grants from the Chang-Gung Medical Research Project (CMRPD no. 33022) and the Taiwan National Science Council (NSC 94-2320-B-182-044, NSC 94-2320-B-182-053) to AMW; and a Taiwan NSC grant 95-3112-B-001-014 to the National Core Facilities for Proteomics, located at the Institute of Biological Chemistry, Academia Sinica.

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Authors and Affiliations

  1. Institute of Biochemical Sciences, National Taiwan University, Taipei, 106, Taiwan

    Shin-Yi Yu & Kay-Hooi Khoo

  2. National Core Facilities for Proteomics and Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei, 115, Taiwan

    Kay-Hooi Khoo

  3. Glyco-Immunochemistry Research Laboratory, Institute of Molecular and Cellular Biology, Chang-Gung University, 333, Kwei-san, Tao-yuan, Taiwan

    Zhangung Yang, Anthony Herp & Albert M. Wu

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  1. Shin-Yi Yu
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Correspondence to Kay-Hooi Khoo or Albert M. Wu.

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Yu, SY., Khoo, KH., Yang, Z. et al. Glycomic mapping of O- and N-linked glycans from major rat sublingual mucin. Glycoconj J 25, 199–212 (2008). https://doi.org/10.1007/s10719-007-9071-y

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  • DOI: https://doi.org/10.1007/s10719-007-9071-y

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