A substantial oxygen isotope effect at O2 in the OMP decarboxylase reaction: mechanistic implications.

Orotidine-5'-monophosphate decarboxylase (OMP decarboxylase, ODCase) catalyzes the decarboxylation of orotidine-5'-monophosphate (OMP) to uridine-5'-monophosphate (UMP). Despite extensive enzymological, structural, and computational studies, the mechanism of ODCase remains incompletely characterized. Herein, carbon kinetic isotope effects were measured for both the natural abundance substrate and a substrate mixture synthesized for the purpose of carrying out the remote double label isotope effect procedure, with O2 of the substrate as the remote position.

Design of LFA-1 antagonists based on a 2,3-dihydro-1H-pyrrolizin-5(7aH)-one scaffold.

A new class of lymphocyte function-associated antigen-1 (LFA-1) antagonists is described. Elaboration of the 2,3-dihydro-1H-pyrrolizin-5(7aH)-one scaffold resulted in the synthesis of potent inhibitors of the LFA-1/ICAM-1 interaction. Along with the in vitro activity, we present the X-ray crystal structure of the complex of compound 9b, in a novel binding mode to the I-domain of LFA-1.

Determination of the absolute configuration and solution conformation of a novel disubstituted pyrrolidine acid A by vibrational circular dichroism.

Compound A, a novel disubstituted pyrrolidine acid, is a member of a new class of agents that are potentially useful for the treatment of diabetes and dyslipidemia. The absolute configuration of this compound was determined by using vibrational circular dichroism (VCD). The results are in agreement with the assignments based on both X-ray analysis and the stereo-selective chemical synthesis.

Theoretical studies of the effect of thio substitution on orotidine monophosphate decarboxylase substrates.

[reaction: see text] The effect of replacing carbonyl oxygens with sulfur in a series of orotidine 5'-monophosphate decarboxylase (ODCase) substrates was studied computationally. Previous experimental results indicate that while 2-thio-orotidine 5'-monophosphate (2-thio-OMP) is a poor substrate for ODCase, 4-thio-orotidine 5'-monophosphate (4-thio-OMP) binds to ODCase, and the resultant k(cat) is measurable. Energetics calculations on 2-thio-1-methyl-orotate and 4-thio-1-methyl-orotate (as models for the 2- and 4-thio-OMPs) indicate that mechanisms involving proton transfer to the 2- or 4-site, regardless of substrate and regardless of whether the 2- or 4-position is a carbonyl or thiocarbonyl, are energetically favorable, as compared to direct decarboxylation without proton transfer.

The gas phase proton affinity of uracil: measuring multiple basic sites and implications for the enzyme mechanism of orotidine 5'-monophosphate decarboxylase.

We have shown for the first time experimentally that the O2 and O4 sites of uracil have different proton affinities, and as implied in previous computational studies, the O4 is more basic and would be energetically preferred in an orotate ribose 5'-monophosphate decarboxylase catalysis mechanism involving proton transfer to oxygen.