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Originally published In Press as doi:10.1074/jbc.M101379200 on April 3, 2001

J. Biol. Chem., Vol. 276, Issue 26, 23221-23229, June 29, 2001
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Isolation and Characterization of Major Glycoproteins of Pigeon Egg White
UBIQUITOUS PRESENCE OF UNIQUE N-GLYCANS CONTAINING Galalpha 1-4Gal*

Noriko SuzukiDagger , Kay-Hooi Khoo§, Hao-Chia Chen, James R. Johnson||, and Yuan C. LeeDagger **

From the Dagger  Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, the § Institute of Biological Chemistry, Academia Sinica, Taipei, 115, Taiwan, the  Endocrinology and Reproduction Research Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, and the || Veterans Affairs Medical Center and Department of Medicine, University of Minnesota, Minneapolis, Minnesota 55417

Ovotransferrin (POT), two ovalbumins (POA(hi) and POA(lo)), and ovomucoid (POM) were isolated from pigeon egg white (PEW). Unlike their chicken egg white counterparts, PEW glycoproteins contain terminal Galalpha 1-4Gal, as evidenced by GS-I lectin (specific for terminal alpha -Gal), anti-P1 (Galalpha 1-4Galbeta 1-4GlcNAcbeta 1-3Galbeta 1-4Glcbeta 1-1Cer) monoclonal antibody, and P fimbriae on uropathogenic Escherichia coli (specific for Galalpha 1-4Gal). Galalpha 1-4Gal on PEW glycoproteins were found in N-glycans releasable by treatment with glycoamidase F. The respective contents of N-glycans in each glycoprotein were 3.5%, POT; 17%, POA(hi); and 31-37%, POM. POA(hi) has four N-glycosylation sites, in contrast to chicken ovalbumin, which has only one. High performance liquid chromatography analysis showed that N-glycans on POA(hi) were highly heterogeneous. Mass spectrometric analysis revealed that the major N-glycans were monosialylated tri-, tetra-, and penta-antennary oligosaccharides containing terminal Galalpha 1-4Gal with or without bisecting N-acetylglucosamine. Oligosaccharide chains terminating in Galalpha 1-4Gal are rare among N-glycans from the mammals and avians that have been studied, and our finding is the first predominant presence of (Galalpha 1-4Gal)-terminated N-glycans.

* This work was supported by National Institutes of Health Research Grant DKO9970 (to N. S. and Y. C. L.), a grant from Academia Sinica (to K.-H. K.), a Department of Veterans Affairs Merit Review (to J. R. J.), and National Institutes of Health Grant DK-47504 (to J. R. J.).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.: 410-516-7041; Fax: 410-516-8716; E-mail: yclee@jhu.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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