Negative homotropic cooperativity and affinity heterogeneity: preparation of yeast glyceraldehyde-3-phosphate dehydrogenase with maximal affinity homogeneity.
- 1 November 1976
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 73 (11) , 3928-3932
- https://doi.org/10.1073/pnas.73.11.3928
Abstract
A three-step procedure including affinity chromatography on NAD+-azobenzamidopropyl-Sepharose has been designed for the purification of yeast glyceraldehyde-3-phosphate dehydrogenase [D-glyceraldehyde-3-phosphate: NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12] with maximized specific activity and maximized homogeneity with respect to affinity for the coenzyme, NAD+.Binding isotherms allow the analysis of cooperativity patterns that disclose both the average ligand affinity in the system and the distribution of ligands among the sites, only for systems with complete affinity homogeneity. The presence of affinity heterogeneity, resulting from multiple oligomeric species differing only in their affinity for coenzyme, gives rise to isotherms which falsely manifest apparent negative cooperativity. A method for distinguishing negative homotropic cooperativity from affinity heterogeneity is suggested.This publication has 28 references indexed in Scilit:
- Specificity of induced conformational changes. Case of yeast glyceraldehyde-3-phosphate dehydrogenaseBiochemistry, 1975
- A simplified method for cyanogen bromide activation of agarose for affinity chromatographyAnalytical Biochemistry, 1974
- Covalent binding of molecules to CNBr-activated agarose: Parameters relevant to the activation and coupling reactionsBiochimica et Biophysica Acta (BBA) - General Subjects, 1974
- Affinity chromatography of nicotinamide–adenine dinucleotide-linked dehydrogenases on immobilized derivatives of the dinucleotideBiochemical Journal, 1973
- Co-operative binding of nicotinamide-adenine dinucleotide to yeast glyceraldehyde-3-phosphate dehydrogenase: I. Equilibrium and temperature-jump studies at pH 8.5 and 40 °CJournal of Molecular Biology, 1971
- Positive and negative cooperativity in yeast glyceraldehyde 3-phosphate dehydrogenaseBiochemistry, 1970
- Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads.1970
- THE BINDING OF NICOTINAMIDE-ADENINE DINUCLEOTIDE TO YEAST D-GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE: TEMPERATURE-JUMP RELAXATION STUDIES ON THE MECHANISM OF AN ALLOSTERIC ENZYMEProceedings of the National Academy of Sciences, 1966
- THE COMPARATIVE ENZYMOLOGY OF LACTIC DEHYDROGENASES. I. PROPERTIES OF THE CRYSTALLINE BEEF AND CHICKEN ENZYMES.1964
- ZINC, A COMPONENT OF YEAST ALCOHOL DEHYDROGENASEProceedings of the National Academy of Sciences, 1955