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N-glycosylation of ovomucin from hen egg white

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Abstract

Ovomucin is a bioactive egg white glycoprotein responsible for the gel properties of fresh egg white and is believed to be involved in egg white thinning, a natural process that occurs during storage. Ovomucin is composed of two subunits: a carbohydrate-rich β-ovomucin with molecular weight of 400–610 KDa and a carbohydrate-poor α-ovomucin with molecular mass of 254 KDa. In addition to limited information on O-linked glycans of ovomucin, there is no study on either the N-glycan structures or the N-glycosylation sites. The purpose of the present study was to characterize the N-glycosylation of ovomucin from fresh eggs using nano LC ESI-MS, MS/MS and MALDI MS. Our results showed the presence of N-linked glycans on both glycoproteins. We found 18 potential N-glycosylation sites in α-ovomucin. 15 sites were glycosylated, one site was found in both glycosylated and non-glycosylated forms and two potential glycosylation sites were found unoccupied. The N-glycans of α-ovomucin found on the glycosylation sites are complex-type structures with bisecting N-acetylglucosamine. MALDI MS of the N-glycans released from α-ovomucin by PNGase F revealed that the most abundant glycan structure is a bisected type of composition GlcNAc6Man3. Two N-glycosylated sites were found in β-ovomucin.

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Abbreviations

GalNAc:

N-acetylgalactosamine

GlcNAc:

N-acetylglucosamine

PTS domain:

mucin domain

MALDI:

matrix-assisted laser desorption/ionization

TOF:

time of flight

MS:

mass spectrometry

LC:

liquid chromatography

ESI:

electro-spray ionization

VWF:

von-Willebrand factor

PNGase F:

peptide N-glycosidase

Hex:

hexose

HexNAc:

N-acetylhexosamine

CK:

cystine knot

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Acknowledgements

This work was supported by grants from Natural Sciences and Engineering Research Council of Canada (NSERC) and Alberta Livestock Meat Agency Inc. (ALMA) to J. Wu.

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  1. Department of Agricultural, Food and Nutritional Science, University of Alberta, 4–10 Ag/For Centre, Edmonton, Alberta, T6G 2P5, Canada

    Marina Offengenden, Messele A. Fentabil & Jianping Wu

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  1. Marina Offengenden
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Correspondence to Jianping Wu.

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Offengenden, M., Fentabil, M.A. & Wu, J. N-glycosylation of ovomucin from hen egg white. Glycoconj J 28, 113–123 (2011). https://doi.org/10.1007/s10719-011-9328-3

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