ABSTRACT
The inactivation kinetics of vacterial glycerol dehydratase (EC 4.2.1.30) in the course of its reaction with adenosylcobalamin (AdoCbl) and its analogs were investigated. It was shown that the inactivation rate of apoenzyme complexes with AdoCbl analogs is determined by the nature of the analogs employed and probably by the rate of their conversion into hydroxycobalamins. A possible inactivation mechanism of glycerol dehydratase is discussed.
Subject(s)
Cobamides/pharmacology , Enterobacter/enzymology , Enterobacteriaceae/enzymology , Hydro-Lyases/antagonists & inhibitors , Apoenzymes/metabolism , Chemical Phenomena , Chemistry , Cobamides/metabolism , Glycerol , Hydro-Lyases/metabolism , Kinetics , Mathematics , Structure-Activity RelationshipABSTRACT
A new method of partial chemical synthesis of adenosylcobalamin (Co alpha-[alpha-5,6-diemethylbenzimidazolyl)]-Co beta-adenosylcobamide, AdoCbl) analogs has been developed. A series of derivatives of AdoCbl modified in the nucleoside and nucleotide ligands and corrin macrocycle have been obtained. The interaction of AdoCl analogs with glycerol dehydratase (EC 4.2.1.30) from Aerobacter aerogenes has been investigated. It has been shown that the nucleoside ligand of AdoCbl provides no essential contribution to the binding of apoenzyme but the preservation of the exact structure of the 1-N and 2-C positions of adenine appears essential for the catalysis. The coordination bond between the Co and nucleotide ligand of AdoCl does not play a decisive role in glycerol dehydratase activity. To form the active site of the glycerol dehydrates, the nucleotide in the AdoCbl structure is essential since nucleotide elimination results in a 100-fold increase of Ki for the corresponding analog. In the binding of AdoCbl with apoenzyme, the main role belongs to the corrin macrocycle, in which the e-propionamide group is significant for binding with apoenzyme, but presumably not essential for catalysis.
