Structural characterization by chromatographic profiling of the oligosaccharides of human immunodeficiency virus (HIV) recombinant envelope glycoprotein gp120 produced in chinese hamster ovary cells

Biomedical Chromatography - Tập 2 Số 6 - Trang 260-270 - 1987
Tsuguo Mizuochi1,2, Michael W. Spellman3, Margot Larkin2, Julie C. Solomon2, Louisette J. Basa3, Ten Feizi2
1On leave of absence from Division of Biomedical Polymer Science, Institute of Comprehensive Medical Science, Fujita-Gakuen Health University School of Medicine, Toyoake Aichi 470–11, Japan.
2Section of Glycoconjugate Research, Medical Research Council, Clinical Research Centre, Watford Road, Harrow, Middlesex HA1 3UJ, UK
3Department of Medicinal and Analytical Chemistry, Genentech Inc., 460 Point San Bruno Boulevard, South San Francisco, California 94080, USA

Tóm tắt

AbstractThis report together with the paper by T. Mizuochi, M. W. Spellman, M. Larkin, J. Solomon, L. J. Basa and T. Feizi (1988) Biochem. J. 254, 599–603 describes the structural elucidation of the N‐linked oligosaccharides of the HIV envelope glycoprotein, gp120 (cloned from the HTLV‐III B isolate and expressed as a secreted fusion protein after transfection of Chinese hamster ovary cells), which is known to bind with high affinity to human T4 lymphocytes. Oligosaccharides were released from peptide by hydrazinolysis, fractionated by paper electrophoresis, high performance lectin affinity chromatography and Bio‐Gel P‐4 column chromatography, and their structures determined by sequential exoglycosidase digestions in conjunction with methylation analysis. The glycoprotein was found to be unique in its diversity of oligosaccharide structures. These include high‐mannose type and hybrid type, as well as four categories of complex type chains: mono‐, bi‐, tri‐ and tetra‐antennary, with or without N‐acetyllactosamine repeats, and with or without a core region fucose residue. Among the sialidase‐treated oligosaccharides no less than 29 structures were identified as follows: magnified image where G=galactose; GN=N‐acetylglucosamine; M=mannose; F=fucose; ±=residues present in a proportion of chains. The actual number of oligosaccharide structures is much greater since before desialylation there was evidence that among the hybrid and complex type chains all but 6% contained sialic acid at the C‐3 position of terminal galactose residues, and partially sialylated forms of the bi‐ and multiantennary chains were present.

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Biomedical Chromatography

Tập 2 Số 6

260-270

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