Further fractionation of the glycoprotein families of porcine zona pellucida by anion-exchange HPLC and some characterization of the separated fractions

Journal of Biochemistry
M NakanoT Tobita

Abstract

The glycoproteins of porcine zonae pellucidae have been fractionated into three families (PZP1-3) by gel filtration HPLC [Nakano et al. (1987) Biochem. Int. 14, 417-423]. However, they still comprise heterogeneous molecular species differing in electric charge. We found that sulfate, but not phosphate, is contained in PZP1-3 by a simple and rapid method for microanalysis of the anionic groups. These families were efficiently separated into many fractions by anion-exchange HPLC. When elution was performed by stepwise increase in NaCl concentration in 8 M urea/20 mM Tris-HCl, pH 8.0, a single distinctive peak emerged for each step. The analyses of amino acids, monosaccharides, and anions of the eight separated fractions of the major family, PZP3, showed that larger amounts of sulfated lactosamine linked to the constituent proteins are present in the fractions that are eluted later: the chain length and/or the chain number of these polylactosamines and the sulfate content increased with stepwise increase in NaCl concentration. Composition analyses also revealed that twice as much N-glycolylneuraminic acid is present as N-acetylneuraminic acid in all fractions. The contents of these sialic acids in the fractions slightly increased ...Continue Reading

Citations

Oct 1, 1992·Molecular Reproduction and Development·E C YurewiczA G Sacco
Jan 28, 1998·Molecular Reproduction and Development·N TanphaichitrD White
Jun 18, 2003·Microscopy Research and Technique·Wei-Hua WangGuang-Ming Wu
Apr 1, 1994·European Journal of Biochemistry·C H HokkeJ F Vliegenthart
Oct 1, 1995·European Journal of Biochemistry·N YonezawaM Nakano
Nov 1, 1995·Andrologia·E Töpfer-PetersenF Sinowatz
Aug 30, 2002·European Journal of Biochemistry·Keiichi IkedaMinoru Nakano
Mar 26, 1999·European Journal of Biochemistry·N YonezawaM Nakano
Dec 4, 2001·Journal of Reproductive Immunology·Liz Howes, Roy Jones
Jul 25, 2003·Biochemical and Biophysical Research Communications·Naoto Yonezawa, Minoru Nakano

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