[Use of immobilization for the study of glyceraldehyde 3-phosphate dehydrogenase. Immobilized dimers of the enzyme].
The immobilized dimers of glyceraldehyde 3-phosphate dehydrogenase have been obtained after dissociation of the tetrameric enzyme molecule linked by one of the subunits with Sepharose 4B. The catalytic parameters (V, Km and pH-dependence of activity) of the immobilized dimers and tetramers of the enzyme are identical. The immobilized dimers are capable of reassociating with the enzyme subunits from solution and possess a higher stability a compared to soluble dehydrogenase. The type of the dimer interaction with the reagents causing the "half-of-the-site reactivity" effect suggests that the immobilized dimers of the yeast and rat skeletal muscle enzymes possess the catalytic activity and, besides, are capable to express cooperative interactions between the active centers of the both subunits. The immobilized dimers of yeast dehydrogenase are able to form hybrids with soluble modified dimers of the yeast enzyme and with those from other sources. A procedure for the enzyme isolation from tissue extracts based on hybridization between the immobilized and soluble dimers of homologous dehydrogenases have been developed.
D000818:Animals; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D009132:Muscles / Q000201:enzymology; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D004800:Enzymes, Immobilized; D046911:Macromolecular Substances; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
Ispol'zovanie immobilizatsii dlia izucheniia glitseral'degid-3-fosfatdegidrogenazy. Immobilizovannye dimery fermenta.