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Glycobiology, 2001, Vol. 11, No. 12 1043-1049
© 2001 Oxford University Press

Structural analysis of sulfated N-linked oligosaccharides in erythropoietin

Nana Kawasaki1,2, Yuji Haishima3, Miyako Ohta2, Satsuki Itoh2, Masashi Hyuga2, Sumiko Hyuga2 and Takao Hayakawa2

2Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501 Japan, and 3Division of Medical Devices, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501 Japan

We previously demonstrated that high-performance liquid chromatography with electrospray ionization mass spectrometry (LC/MS) equipped with a graphitized carbon column (GCC) is useful for the structural analysis of carbohydrates in glycoproteins. Using LC/MS with GCC, sulfated N-linked oligosaccharides were found in erythropoietin (EPO) expressed in baby hamster kidney cells. Sulfation occurs in a part of the N-linked oligosaccharides in the EPO. Sulfated monosaccharide residue in the sulfated N-linked oligosaccharide was determined by exoglycosidase digestion followed by sugar mapping by LC/MS. The linkage position and branch-location of the sulfate group in the tetraantennary oligosaccharide were analyzed by 1H-nuclear magnetic resonance. It was suggested that sulfation occurs on the C-6 position of GlcNAc located in the GlcNAcß1-4Man{alpha}1-3 branch.

1 To whom correspondence should be addressed


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