Unknown

Dataset Information

0

Isopentenyl diphosphate isomerase catalyzed reactions in D2O: product release limits the rate of this sluggish enzyme-catalyzed reaction.


ABSTRACT: The E. coli isopentenyl diphosphate isomerase (IDI) catalyzed reaction of isopentenyl diphosphate (IPP) in D(2)O gives a 66% yield of dimethylallyl diphosphate labeled with deuterium at the (E)-methyl group (d-DMAPP) and a 34% yield of IPP labeled with 1 mol of deuterium at C-2 (d-IPP). This shows that the release to D(2)O of the initial product of the IDI-catalyzed reaction (d-DMAPP) is slower than its conversion to d-IPP. Product dissociation is therefore rate determining for isomerization of IPP with a rate constant k(dis) ? k(cat) = 0.08 s(-1). The data provide an estimated rate constant of k(as) = 6 × 10(3) M(-1) s(-1) for binding of DMAPP to E. coli IDI that is similar to rate constants determined for the binding of N-protonated 2-amino ethyl diphosphate intermediate analogs to IDI from yeast [Reardon, J. E.; Abeles, R. H. Biochemistry1986, 25, 5609-5616]. We propose that ligand binding to IDI is relatively slow because there is a significant kinetic barrier to reorganization of the initial encounter complex between enzyme, substrate, and an essential Mg(2+) to form the Michaelis complex where the metal cation bridges the protein and the substrate diphosphate group.

SUBMITTER: Jonnalagadda V 

PROVIDER: S-EPMC3334862 | biostudies-literature | 2012 Apr

REPOSITORIES: biostudies-literature

altmetric image

Publications

Isopentenyl diphosphate isomerase catalyzed reactions in D2O: product release limits the rate of this sluggish enzyme-catalyzed reaction.

Jonnalagadda Venkatadurga V   Toth Krisztina K   Richard John P JP  

Journal of the American Chemical Society 20120405 15


The E. coli isopentenyl diphosphate isomerase (IDI) catalyzed reaction of isopentenyl diphosphate (IPP) in D(2)O gives a 66% yield of dimethylallyl diphosphate labeled with deuterium at the (E)-methyl group (d-DMAPP) and a 34% yield of IPP labeled with 1 mol of deuterium at C-2 (d-IPP). This shows that the release to D(2)O of the initial product of the IDI-catalyzed reaction (d-DMAPP) is slower than its conversion to d-IPP. Product dissociation is therefore rate determining for isomerization of  ...[more]

Similar Datasets

| S-EPMC145486 | biostudies-literature
| S-EPMC2528281 | biostudies-literature
| S-EPMC3237117 | biostudies-literature
| S-EPMC2912430 | biostudies-literature
| S-EPMC2494526 | biostudies-literature
| S-EPMC2374249 | biostudies-literature
| S-EPMC3079984 | biostudies-literature
| S-EPMC5650523 | biostudies-literature
| S-EPMC3438293 | biostudies-literature