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J Biol Chem, Vol. 275, Issue 11, 7731-7742, March 17, 2000

Structural Analysis of Murine Zona Pellucida Glycans
EVIDENCE FOR THE EXPRESSION OF CORE 2-TYPE O-GLYCANS AND THE Sd^a ANTIGEN^*

Richard L. Easton, Manish S. Patankar^§, Frank A. Lattanzio^§, Trey H. Leaven^§, Howard R. Morris^¶, Gary F. Clark^§¶, and Anne Dell^¶

From the  Department of Biochemistry, Imperial College of Science, Technology and Medicine, London SW7 2AY, United Kingdom and ^§ Department of Physiological Sciences, Eastern Virginia Medical School, Norfolk, Virginia 23501

Murine sperm initiate fertilization by binding to specific oligosaccharides linked to the zona pellucida, the specialized matrix coating the egg. Biophysical analyses have revealed the presence of both high mannose and complex-type N-glycans in murine zona pellucida. The predominant high mannose-type glycan had the composition Man₅GlcNAc₂, but larger oligosaccharides of this type were also detected. Biantennary, triantennary, and tetraantennary complex-type N-glycans were found to be terminated with the following antennae: Gal1-4GlcNAc, NeuAc2-3Gal1-4GlcNAc, NeuGc2-3Gal1-4GlcNAc, the Sd^a antigen (NeuAc2-3[GalNAc1-4]Gal1-4GlcNAc, NeuGc2-3[GalNAc1-4]Gal1-4GlcNAc), and terminal GlcNAc. Polylactosamine-type sequence was also detected on a subset of the antennae. Analysis of the O-glycans indicated that the majority were core 2-type (Gal1-4GlcNAc1-6[Gal1-3]GalNAc). The 1-6-linked branches attached to these O-glycans were terminated with the same sequences as the N-glycans, except for terminal GlcNAc. Glycans bearing Gal1-4GlcNAc1-6 branches have previously been suggested to mediate initial murine gamete binding. Oligosaccharides terminated with GalNAc1-4Gal have been implicated in the secondary binding interaction that occurs following the acrosome reaction. The significant implications of these observations are discussed.

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^* This work was supported by a grant from the Biotechnology and Biological Sciences Research Council (to H. R. M. and A. D.) and a grant from the Wellcome Trust (to H. R. M. and A. D.). This study was also supported by the Jeffress Trust (to G. F. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Tel.: 44 171 594 5219; Fax: 44 171 225 0458: E-mail: a.dell@ic.ac.uk (A. Dell and H. R. Morris) or Tel.: 757 446 5650; Fax: 757 624 2270; E-mail: gfc@ borg.evms.edu (G. F. Clark).

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Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.

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