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Structural and Quantitative Characterization of Mucin-Type O-Glycans and the Identification of O-Glycosylation Sites in Bovine Submaxillary Mucin.


ABSTRACT: Bovine submaxillary mucin (BSM) is a gel-forming glycoprotein polymer, and Ser/Thr-linked glycans (O-glycans) are important in regulating BSM's viscoelasticity and polymerization. However, details of O-glycosylation have not been reported. This study investigates the structural and quantitative characteristics of O-glycans and identifies O-glycosylation sites in BSM using liquid chromatography-tandem mass spectrometry. The O-glycans (consisting of di- to octa-saccharides) and their quantities (%) relative to total O-glycans (100%; 1.1 pmol per 1 ?g of BSM) were identified with 14 major (>1.0%), 12 minor (0.1%-1.0%), and eight trace (<0.1%) O-glycans, which were characterized based on their constituents (sialylation (14 O-glycans; 81.9%, sum of relative quantities of each glycan), non-sialylation (20; 18.1%), fucosylation (20; 17.5%), and terminal-galactosylation (6; 3.6%)) and six core structure types [Gal-GalNAc, Gal-(GlcNAc)GalNAc, GlcNAc-GalNAc, GlcNAc-(GlcNAc)GalNAc, and GalNAc-GalNAc]. O-glycosylation sites were identified using O-glycopeptides (bold underlined; 56SGETRTSVI, 259SHSSSGRSRTI, 272GSPSSVSSAEQI, 307RPSYGAL, 625QTLGPL, 728TMTTRTSVVV, and 1080RPEDNTAVA) obtained from proteolytic BSM; these sites are in the four domains of BSM. The gel-forming mucins share common domain structures and glycosylation patterns; these results could provide useful information on mucin-type O-glycans. This is the first study to characterize O-glycans and identify O-glycosylation sites in BSM.

SUBMITTER: Kim J 

PROVIDER: S-EPMC7226346 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Structural and Quantitative Characterization of Mucin-Type <i>O</i>-Glycans and the Identification of <i>O</i>-Glycosylation Sites in Bovine Submaxillary Mucin.

Kim Jihye J   Ryu Changsoo C   Ha Jongkwan J   Lee Junmyoung J   Kim Donghwi D   Ji Minkyoo M   Park Chi Soo CS   Lee Jaeryong J   Kim Dae Kyong DK   Kim Ha Hyung HH  

Biomolecules 20200420 4


Bovine submaxillary mucin (BSM) is a gel-forming glycoprotein polymer, and Ser/Thr-linked glycans (<i>O</i>-glycans) are important in regulating BSM's viscoelasticity and polymerization. However, details of <i>O</i>-glycosylation have not been reported. This study investigates the structural and quantitative characteristics of <i>O</i>-glycans and identifies <i>O</i>-glycosylation sites in BSM using liquid chromatography-tandem mass spectrometry. The <i>O</i>-glycans (consisting of di- to octa-s  ...[more]

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