Structural study of the sugar chains of CD36 purified from bovine mammary epithelial cells: occurrence of novel hybrid-type sugar chains containing the Neu5Ac alpha 2-->6GalNAc beta 1-->4GlcNAc and the Man alpha 1-->2Man alpha 1-->3Man alpha 1-->6Man groups.
CD36 is a glycoprotein included in the bovine milk fat globule membrane derived from mammary secretory epithelial cells during lactation. Asparagine-linked sugar chains were quantitatively released from CD36 as oligosaccharides by hydrazinolysis. These sugar chains were converted to radioactive oligosaccharides by reduction with NaB3H4 and separated into neutral and acidic fractions by paper electrophoresis. Most of the acidic oligosaccharides were converted to neutral ones by sialidase digestion, indicating that they are sialyl derivatives. The neutral and sialidase-treated acidic oligosaccharides were fractionated by Bio-Gel P-4 column chromatography in combination with serial chromatography on immobilized lectin columns including a Wistaria floribunda agglutinin (WFA)-agarose column. WFA is known to bind oligosaccharides terminating with either an alpha- or beta-N-acetylgalactosamine residue. Structural studies of oligosaccharides in each fraction by sequential exoglycosidase digestion as well as methylation analysis revealed that CD36 contains high mannose-type, hybrid-type, and bi, tri-, and tetraantennary complex-type sugar chains. A portion of the hybrid-type and the complex-type sugar chains which bound to a WFA-agarose column (28% of all oligosaccharides) contained the GalNAc beta 1-->4GlcNAc group(s) instead of the Gal beta 1-->4GlcNAc group(s) in their outer chain moieties. Like oligosaccharides found in human luteinizing hormone [Weisshaar, G., Hiyama, J., Renwick, A. G., & Nimtz, M. (1991) Eur. J. Biochem. 195, 257-268], some of the GalNAc beta 1-->4GlcNAc groups found in the CD36 oligosaccharides were sialylated as the Neu5Ac alpha 2-->6GalNAc group. Furthermore, most of the hybrid-type sugar chains of CD36 with the Gal/GalNAc beta 1-->4GlcNAc beta 1-->2 outer chain on their Man alpha 1-->3 arm contained an unusual Man alpha 1-->2Man alpha 1-->3 group on their Man alpha 1-->6 arm.
Nakata
N
N
Department of Biochemistry, University of Tokyo.
Furukawa
K
K
Greenwalt
D E
DE
Sato
T
T
Kobata
A
A
D000818:Animals; D015703:Antigens, CD / Q000737:chemistry* / Q000302:isolation & purification; D001216:Asparagine; D018955:CD36 Antigens; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002846:Chromatography, Affinity; D004848:Epithelium / Q000737:chemistry; D005260:Female; D006023:Glycoproteins / Q000737:chemistry*; D006026:Glycoside Hydrolases; D007202:Indicators and Reagents; D007425:Intracellular Membranes / Q000737:chemistry; D037102:Lectins; D008321:Mammary Glands, Animal / Q000737:chemistry*; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification
D015703:Antigens, CD; D018955:CD36 Antigens; D006023:Glycoproteins; D007202:Indicators and Reagents; D037102:Lectins; D009844:Oligosaccharides; D001216:Asparagine; D006026:Glycoside Hydrolases