L. Reynolds, G. Garcia, J. Kozarich
Jul 26, 1988
Citations
2
Influential Citations
21
Citations
Quality indicators
Journal
Biochemistry
Abstract
A series of [p-(halomethyl)benzoyl]formates have been investigated as substrates for benzoylformate decarboxylase. These analogues vary from acting as normal substrates to acting as potent competitive inhibitors. The fluoro analogue is a substrate with Km (190 microM) and turnover number (20 s-1) similar to those of benzoylformate (Km = 340 microM; 81 s-1). The bromo analogue is a competitive inhibitor (Ki = 0.3 microM) and exhibits processing to eliminate bromide and form (p-methylbenzoyl)thiamin pyrophosphate. This modified cofactor hydrolyzes to form the p-methylbenzoate in quantitative yield. The chloro analogue [Km(app) = 21 microM] partitions between these two pathways such that 0.6% of the analogue ultimately forms p-methylbenzoate. These data are consistent with the interpretation that the leaving group potential of the halogen determines the enzymic fate of the analogue and that the potent inhibition observed for the bromo analogue is due to covalent modification of the cofactor.