[The effect of various acylating agents on the catalytic properties and structure of aspartate aminotransferase].
Changes of quaternary structure and conformation of molecule concomitant with inactivation were observed in the course of aspartate transaminase acylation by maleic, citraconic, dimethylmaleic and succinic anhydrides. It was established that acylation of 10-12 xi-amino groups of lysine did not induce the dissociation of transaminase into subunits. Further acylation of amino groups (2 groups if dimethylmaleic anhydrade was used as acylating agent) induced dissociation of transaminase dimer into subunits. These data were obtained by sedimentation analysis. The dissociation was accompanied with a sharp decrease of correlation time (from 18 nsec to 9 nsec) of the paramagnetic label covalently bound to the protein. The obtained results allow us to distinguish three types of xi-aminogroups of aspartate transaminase: exposed (about 12 residues), "contact" (2 residues) located in the vicinity to complementary surfaces of subunits and buried (about 6 residues). The stepwise  inactivation occurred during the acylation as a result of conformational changes or appearance of sterical hindrances in the cataytic site of the enzyme. The thiol groups were not modified in transaminase molecule under experimental conditions used. Aspartate transaminase treated with citraconic or dimethylmaleic anhydride may be deacylated under mild conditions. After reacylation the quaternary structure was reconstituted and catalytic activity was almost fully restored.
D000215:Acylation; D000812:Anhydrides / Q000494:pharmacology; D001219:Aspartate Aminotransferases / Q000378:metabolism; D001665:Binding Sites; D002264:Carboxylic Acids / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D046911:Macromolecular Substances; D008298:Maleates / Q000494:pharmacology; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D013386:Succinates / Q000494:pharmacology
D000812:Anhydrides; D002264:Carboxylic Acids; D046911:Macromolecular Substances; D008298:Maleates; D013386:Succinates; D001219:Aspartate Aminotransferases
Bliianie razlichnykh atsiliruiushchikh agentov na kataliticheskie svoĭstva i strukturu aspartat-aminotransferazy