PMID: 6405788May 30, 1983Paper

Use of concanavalin A as a topographical probe for protein-protein interaction. Application to lactose synthase

Biochimica Et Biophysica Acta
S S WongT K Lee

Abstract

Galactosyltransferase (EC 2.4.1.38) has been shown to bind to Con A-Sepharose. Concentrations of methyl-alpha-mannoside greater than 0.7 M were required to release the enzyme from the immobilized lectin. Molecular weight determination by gel filtration revealed that galactosyltransferase formed a 1:1 complex with concanavalin A. Preincubation of the enzyme with excess concanavalin A did not affect its catalytic activity either in the presence or absence of alpha-lactalbumin. The galactosyltransferase-concanavalin A complex was retained on an alpha-lactalbumin-Sepharose column in the presence of N-acetylglucosamine and manganese chloride and was eluted from the column in their absence. Galactosyltransferase immobilized onto a Con A-Sepharose was still active either in the presence or absence of alpha-lactalbumin. Lactose synthase activity was also observed when the galactosyltransferase-concanavalin A complex was assayed with alpha-lactalbumin immobilized on Sepharose. These data indicate that the carbohydrate moiety of galactosyltransferase is involved in neither the catalytic process nor the binding of alpha-lactalbumin and must be linked to the enzyme at a location where it does not present any steric hindrance on the binding...Continue Reading

References

Aug 1, 1977·FEBS Letters·G P RoyerK Aso
Mar 29, 1977·Biochimica Et Biophysica Acta·C R GerenK E Ebner
Jan 1, 1976·Archives of Biochemistry and Biophysics·C R GerenK E Ebner
May 15, 1974·European Journal of Biochemistry·B S KhatraK Brew
Mar 1, 1971·The Biochemical Journal·A E Clarke, M A Denborough
Feb 1, 1968·Proceedings of the National Academy of Sciences of the United States of America·K BrewR L Hill
Jun 1, 1981·The Biochemical Journal·K YamamotoT Osawa
Jan 4, 1965·Biochimica Et Biophysica Acta·I J GOLDSTEINJ M MERRICK

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Citations

Jan 1, 1984·The International Journal of Biochemistry·L J Wong, S S Wong
Jan 1, 1989·Critical Reviews in Biochemistry and Molecular Biology·M J Kronman
Dec 20, 1985·Biochimica Et Biophysica Acta·D R JohnsonS S Wong

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