19F-n.m.r. studies of 3',5'-difluoromethotrexate binding to Lactobacillus casei dihydrofolate reductase. Molecular motion and coenzyme-induced conformational changes.

  title={19F-n.m.r. studies of 3',5'-difluoromethotrexate binding to Lactobacillus casei dihydrofolate reductase. Molecular motion and coenzyme-induced conformational changes.},
  author={G. Marius Clore and Angela M. Gronenborn and Berry Birdsall and James Feeney and Gordon C. K. Roberts},
  journal={The Biochemical journal},
  volume={217 3},
19F-n.m.r. spectroscopy was used to study the binding of 3',5'-difluoromethotrexate to dihydrofolate reductase (tetrahydrofolate dehydrogenase) from Lactobacillus casei. The benzoyl ring of the bound difluoromethotrexate was found to 'flip' about its symmetry axis, and the rate (7.3 X 10(3) s-1 at 298 K) and activation parameters for this process were determined by lineshape analysis of the 19F-n.m.r. spectrum at a series of temperatures in the range 273-308 K. The contributions to the barrier… 
23 Citations
Trimethoprim binding to Lactobacillus casei dihydrofolate reductase: a 13C NMR study using selectively 13C-enriched trimethoprim.
The large shift observed for C6 in all complexes indicates that the basic folded conformation is present in all of them, and that the pyrimidine ring is protonated at N1 in all the complexes of trimethoprim with the enzyme and coenzymes.
Structural comparisons of complexes of methotrexate analogues with Lactobacillus casei dihydrofolate reductase by two-dimensional 1H NMR at 500 MHz.
In the complexes formed with the dihalomethotrexate analogues, the glutamic acid and pteridine ring moieties were shown to bind to the enzyme in a manner similar to that found in the methotrexates-enzyme complex.
Mobility of the spin-labeled side chains of some novel antifolate inhibitors in their complexes with dihydrofolate reductase.
Four spin-labeled inhibitors of dihydrofolate reductase (DHFR) have been synthesized and the spectra indicate that when these inhibitors are bound to the enzyme the TEMPO group is highly immobilized with correlation time, tau c, 4-20ns.
Multinuclear NMR characterization of two coexisting conformational states of the Lactobacillus casei dihydrofolate reductase-trimethoprim-NADP+ complex.
The complex of Lactobacillus casei dihydrofolate reductase with trimethoprim and NADP+ exists in solution as a mixture of approximately equal amounts of two slowly interconverting conformational states, and a partial structural model has been proposed.
Structure and dynamics in solution of the complex of lactobacillus casei dihydrofolate reductase with the new lipophilic antifolate drug trimetrexate
The binding site for trimetrexate is well defined and this was compared with the binding sites in related complexes formed with methotrexate and trimethoprim, and no major conformational differences were detected between the different complexes.
NMR Studies Of complexes of L. Casei Dihydrofolate Reductase with Antifolate Drugs: Multiple Conformations and Conformational Selection of Bound Rotational Isomers
The enzyme is of considerable pharmacological interest being the target for several clinically useful ‘antifolate’ drugs such as methotrexate (anti-neoplastic), trimethoprim ( anti-bacterial) and pyrimethamine (Anti-malarial) 3.
NMR Studies of Protein-Ligand Interactions: Dihydrofolate Reductase
The study of protein-ligand interactions has proved to be one of the more fruitful applications of high-resolution nmr to biological problems. The specific recognition of small molecules by proteins
Fluorine NMR of proteins