[Immobilization of glyceraldehyde-3-phosphate dehydrogenase by non-covalent binding to specific antibodies and Fab-fragments coupled to Sepharose].
Rabbit antibodies to rat skeletal muscle glyceraldehyde-3-phosphate dehydrogenase, as well as monovalent Fab fragments of these antibodies were coupled to CNBr-activated Sepharose 4B. Rat skeletal muscle glyceraldehyde-3-phosphate dehydrogenase was then immobilized on a matrix by non-covalent binding to specific antibodies. Immobilized enzyme retains approximately 90% catalytic activity of the soluble dehydrogenase; pH optimum of activity and the Km value observed are changed as compared to the enzyme in solution. Glyceraldehyde-3-phosphate dehydrogenase immobilized on specific antibodies is shown to undergo adenine nucleotide-induced dissociation into dimers. The immobilized dimeric form of the enzyme thus obtained is catalytically active and capable of reassociating with the dimers of apoglyceraldehyde-3-phosphate dehydrogenase added in solution to the suspension of Sepharose.
D000906:Antibodies; D001051:Apoenzymes; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000276:immunology / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007140:Immunoglobulin Fab Fragments; D007700:Kinetics; D046911:Macromolecular Substances; D009132:Muscles / Q000201:enzymology; D012685:Sepharose
D000906:Antibodies; D001051:Apoenzymes; D004800:Enzymes, Immobilized; D007140:Immunoglobulin Fab Fragments; D046911:Macromolecular Substances; D012685:Sepharose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
Nekovalentnaia immobilizatsiia glitseral'degid-3-fosfatdegidrogenazy na antitelakh i Fab-fragentakh antitel, sviazannykh s sefarozoĭ.