Immunochemical studies of infectious mononucleosis. V. Isolation and characterization of a glycoprotein from goat erythrocyte membranes

M. A. Fletcher, T. M. Lo, W. R. Graves

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14 Scopus citations

Abstract

A glycoprotein was isolated from goat erythrocyte membranes by extraction with hot 75% ethanol. The glycoprotein was purified by ethanol precipitation, phosphocellulose chromatography gel filtration, ethyl : ether and chloroform : methanol extraction. In aqueous phosphate buffered saline, pH 7, the glycoprotein was in an aggregated state with a sedimentation coefficient (S(obs)) of 21. Addition of 0.5% sodium dodecyl sulphate (SDS) lowered S(obs) to 1.5. Electrophoresis of the glycoprotein on polyacrylamide gels containing phosphate buffered 0.1% SDS gave a single band, staining with both periodic acid Schiff (PAS) and Coomassie Blue (CB). The apparent m.w., calculated from retardation coefficient, was 25,000. Electrophoresis of the glycoprotein on 1% SDS gels buffered with Tris acetate (pH 7.4) showed a major band of similar (23,500) apparent m.w. plus four other PAS and CB staining bands of lower mobility. With 131I labeled glycoprotein, recovery of bands from gels, sialic acid analysis, heterophile antigen activity, and re electrophoresis, it was shown that these additional bands were aggregated forms of a single or closely related glycoprotein species. The purified glycoprotein contained 50% carbohydrate with molar ratios of sialic acid : galactose : mannose : galactosamine : glucosamine of 3.1:2.1:0.1:1.6:1. The glycoprotein was highly reactive with the Paul Bunnel heterophile antibody in the sera of patients with infectious mononucleosis, with Limulus polyphemus lectin and weakly reactive with wheat germ agglutinin. These reactivities were destroyed by neuraminidase treatment or by alkaline sodium borohydride. The native glycoprotein did not react with lectins from Canavalia enisformis, Phaseolus vulgaris, Ricinus communis, or Vicia graminea although it was reactive with the latter two after neuraminidase treatment.

Original languageEnglish (US)
Pages (from-to)722-729
Number of pages8
JournalJournal of Immunology
Volume117
Issue number3
StatePublished - Dec 1 1976

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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