title: Subunit interaction in mammalian aldolases.
abstract: Enzyme inactivation was utilized to study subunit interaction in the homotetrameric glycolytic enzyme, aldolase. Isoenzymes from rabbit liver and skeletal muscle were inactivated in the presence of Pi and d-glyceraldehyde-P to a maximum stoichiometry of one modification per aldolase subunit. Subunit modification increased net negative charge on each subunit surface and was used to resolve modified aldolase isoenzymes into various chromatographic species. A combination of anion-(Mono Q) and cation- (Mono S) exchange chromatography separated the modified aldolase homotetramers into three distinct enzyme populations: unchanged enzyme, fully modified enzyme corresponding to one ligand molecule incorporated per subunit and partially modified enzyme in which only one subunit out of four is modified. Both fully and partially modified species were devoid of catalytic activity. Activity loss through modification of a single subunit in both aldolase isoenzymes indicates tightly coupled communication between subunit active sites and suggests simple functional regulation of aldolases.
authors.affiliation: Departement de Biochimie, Faculté de Medécine, Université de Montréal, CP 6128, Station Centre-Ville Montréal, Québec, H3C 3J7 Canada.
authors.forename: J
authors.initials: J
authors.lastname: Sygusch
authors.forename: D
authors.initials: D
authors.lastname: Beaudry
mesh_terms: D000818:Animals; D002852:Chromatography, Ion Exchange; D004789:Enzyme Activation; D005634:Fructose-Bisphosphate Aldolase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D005986:Glyceraldehyde 3-Phosphate / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D008024:Ligands; D008099:Liver / Q000201:enzymology; D008322:Mammals / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000201:enzymology; D010710:Phosphates / Q000494:pharmacology; D011487:Protein Conformation; D011817:Rabbits
chemical_list: D007527:Isoenzymes; D008024:Ligands; D010710:Phosphates; D005986:Glyceraldehyde 3-Phosphate; D005634:Fructose-Bisphosphate Aldolase