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Glycobiology, 2001, Vol. 11, No. 3 195-208
© 2001 Oxford University Press

N-linked oligosaccharides of cobra venom factor contain novel {alpha}(1-3)galactosylated Lex structures

D. Channe Gowda1,5, John Glushka6, Herman van Halbeek2,6, Rao N. Thotakura7, Reinhard Bredehorst3,5 and Carl-Wilhelm Vogel4,5

5Department of Biochemistry and Molecular Biology, and Vincent T. Lombardi Cancer Center, Georgetown University Medical Center, 3900 Reservoir Road, NW, Washington, DC 20007, USA, 6Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA, and 7Molecular and Cellular Endocrinology Branch, NIDDK, National Institutes of Health, Bethesda, MD 20892, USA

Cobra venom factor (CVF), a nontoxic, complement-activating glycoprotein in cobra venom, is a functional analog of mammalian complement component C3b. The carbohydrate moiety of CVF consists exclusively of N-linked oligosaccharides with terminal {alpha}1-3-linked galactosyl residues, which are antigenic in human. CVF has potential for several medical applications, including targeted cell killing and complement depletion. Here, we report a detailed structural analysis of the oligosaccharides of CVF. The structures of the oligosaccharides were determined by lectin affinity chromatography, antibody affinity blotting, compositional and methylation analyses, and high-resolution 1H-NMR spectroscopy. Approximately 80% of the oligosaccharides are diantennary complex-type, ~12% are tri- and tetra-antennary complex-type, and ~8% are oligomannose type structures. The majority of the complex-type oligosaccharides terminate in Gal{alpha}1-3Galß1-4(Fuc{alpha}1-3)GlcNAcß1, a unique carbohydrate structural feature abundantly present in the glycoproteins of cobra venom.

1 To whom correspondence should be addressed

2 Present address: Glycobiology Research and Training Center, School of Medicine, University of California at San Diego, La Jolla, CA 92093-0687, USA.

3 Present address: Department of Biochemistry and Molecular Biology, University of Hamburg, Hamburg 20146, Germany.

4 Present address: Cancer Research Center of Hawaii, University of Hawaii, 1236 Lauhala Street, Honolulu, HI 96813, USA.


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