Ch. Wei et al., COMPARISON OF BRANCHED-CHAIN AND TIGHTLY COUPLED REACTION-MECHANISMS FOR PROSTAGLANDIN-H SYNTHASE, Biochemistry, 34(26), 1995, pp. 8499-8512
Two types of mechanisms have been proposed to account for the combinat
ion of peroxidase and cyclooxygenase activities in prostaglandin H syn
thase (PGHS). One, a branched-chain mechanism [Dietz, R., et al. (1988
) Eur. J. Biochem. 171, 321-328], postulates that the cyclooxygenase r
eaction propagates essentially independently of peroxidase catalysis.
The second, a tightly coupled mechanism [Bakovic, M., & Dunford, H. B.
(1994) Biochemistry 33, 6475-6482], postulates that peroxidase cataly
sis is an integral part of cyclooxygenase propagation. Qualitative and
quantitative predictions from the two mechanisms have been compared w
ith several observed characteristics of the PGHS reaction with arachid
onate, including the ability to accumulate PGG(2) and oxidized enzyme
intermediates, the stoichiometry between cosubstrate and fatty acid co
nsumption, and the hydroperoxide activator requirement. The observed c
haracteristics, particularly the accumulation of micromolar levels of
PGG(2) even in the presence of cosubstrate and the stoichiometry betwe
en cosubstrate oxidation and fatty acid oxygenation of less than 1.3 (
compared to a theoretical maximum of 2.0), were largely consistent wit
h predictions from the branched-chain mechanism, but contradicted impo
rtant predictions of the tightly coupled mechanism. These results indi
cate that PGHS catalysis is more accurately described by the branched-
chain mechanism than by the tightly coupled mechanism.