Isolation and characterization of major glycoproteins of pigeon egg white: ubiquitous presence of unique N-glycans containing Galalpha1-4Gal.
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 Galalpha1-4Gal, as evidenced by GS-I lectin (specific for terminal alpha-Gal), anti-P(1) (Galalpha1-4Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glcbeta1-1Cer) monoclonal antibody, and P fimbriae on uropathogenic Escherichia coli (specific for Galalpha1-4Gal). Galalpha1-4Gal 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 Galalpha1-4Gal with or without bisecting N-acetylglucosamine. Oligosaccharide chains terminating in Galalpha1-4Gal are rare among N-glycans from the mammals and avians that have been studied, and our finding is the first predominant presence of (Galalpha1-4Gal)-terminated N-glycans.
Suzuki
N
N
Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
Khoo
K H
KH
Chen
H C
HC
Johnson
J R
JR
Lee
Y C
YC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002240:Carbohydrate Sequence; D010856:Columbidae; D003207:Conalbumin / Q000737:chemistry / Q000302:isolation & purification; D004187:Disaccharides / Q000032:analysis* / Q000276:immunology; D004527:Egg Proteins / Q000737:chemistry* / Q000302:isolation & purification; D004529:Egg White / Q000032:analysis*; D005690:Galactose / Q000737:chemistry; D005697:Galactosides / Q000032:analysis / Q000276:immunology; D006031:Glycosylation; D006386:Hemagglutination Tests; D037102:Lectins / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D010047:Ovalbumin / Q000737:chemistry / Q000302:isolation & purification; D010059:Ovomucin / Q000737:chemistry / Q000302:isolation & purification; D037121:Plant Lectins; D011134:Polysaccharides / Q000737:chemistry* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D004187:Disaccharides; D004527:Egg Proteins; D005697:Galactosides; C045475:Griffonia simplicifolia lectins; D037102:Lectins; D009005:Monosaccharides; D037121:Plant Lectins; D011134:Polysaccharides; D003207:Conalbumin; D010059:Ovomucin; C056853:6-O-galactopyranosylgalactose; D010047:Ovalbumin; D005690:Galactose