Isoproterenol inhibition of horse serum cholinesterase is connected with subunit dissociation.
Tetrameric cholinesterase from horse serum undergoes concentration-dependent dissociation. The dimer is highly stable so that even on SDS polyacrylamide gels subunit dissociation to the 80-kDa polypeptide chains is incomplete. Glutaraldehyde cross-linking confirms this finding, giving rise to a tetramer: dimer ratio of approximately 1:1. The beta-adrenergic agent isoproterenol acts as an inhibitor of the enzyme with respect to butyrylthiocholine hydrolysis; inhibition kinetics point to a dissociative effect of the ligand as the underlying mechanism (Söylemez, Z. & Ozer, I. (1985) Comp. Biochem. Physiol. 81c, 433-437). Evidence from sedimentation analysis confirms this hypothetical mechanism: the sedimentation coefficient in the presence of saturating concentrations of both the substrate butyrylthiocholine and the inhibitor isoproterenol shows a 35 +/- 5% decrease; in high speed sedimentation equilibria the weight average molecular mass is shifted from the tetramer (Mr = 312 +/- 12 kDa) to the dimer (Mr = 160 +/- 10 kDa). The transition is complete at isoproterenol concentrations below saturation. Applying glutaraldehyde cross-linking to monitor the particle distribution at varying isoproterenol concentrations confirms the change in quaternary structure in a qualitative way. Enzyme concentrations applied in the present experiments are in the range of the concentration of cholinesterase in horse serum. Therefore the dissociative mechanism of isoproterenol on the enzyme may be of biological significance.
D000818:Animals; D002800:Cholinesterase Inhibitors; D002802:Cholinesterases / Q000097:blood*; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005976:Glutaral / Q000032:analysis; D006736:Horses; D007545:Isoproterenol / Q000494:pharmacology*; D046911:Macromolecular Substances; D008970:Molecular Weight
D002800:Cholinesterase Inhibitors; D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D002802:Cholinesterases; D007545:Isoproterenol; D005976:Glutaral