Abstract
Human seminal transferrin (HSmT) is an iron-containing glycoprotein whose structural properties have not been adequately investigated. The carbohydrate content of the purified glycoprotein amount to 6.1%, and monosaccharide analysis revealed the major oligosaccharide moiety to be of the N-glycoside type. The carbohydrate chains were released from the iron-free form by digestion with peptide N-glycosidase F (PNGase F) in the presence of detergents such as SDS andΒ-octylglucoside. After ethanol precipitation and fractionation on Bio-Gel P-6 and Bio-Gel P-2, the oligosaccharide was further purified on Mono-Q and desalted on Bio-Gel P-2. By 600-MHz1H-NMR spectroscopy, the primary structure of the major N-linked oligosaccharide component was established to be:
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Abbreviations
- HSmT:
-
human seminal transferrin
- HSrT:
-
human serum transferrin
- PNGase F:
-
peptide-N4-(N-acetyl-Β-glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52), commonly known as peptide N-glycosidase F
- SDS-PAGE:
-
sodium dodecyl sulphate-polyacrylamide gel electrophoresis
- GLC:
-
gas-liquid chromatography
- FPLC:
-
fast liquid protein chromatography
- EDTA:
-
ethylenediaminetetraacetic acid, disodium salt
- PMFS:
-
phenylmethylsulfonyl fluoride
- GlcNAc:
-
N-acetylglucosamine
- NeuAc:
-
N-acetylneuraminic acid
- Man, Gal:
-
galactose
- Fuc:
-
fucose
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D'Andrea, G., D'Alessandro, A.M., Salucci, M.L. et al. Primary structure of the major glycan from human seminal transferrin. J Protein Chem 13, 31–36 (1994). https://doi.org/10.1007/BF01891990
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DOI: https://doi.org/10.1007/BF01891990