Interpretation of nuclear magnetic resonance spectra for Lactobacillus casei dihydrofolate reductase based on the X-ray structure of the enzyme-methotrexate-NADPH complex.
The three-dimensional molecular structure of Lactobacillus casei dihydrofolate reductase complexed with NADPH and methotrexate has been used to interpret published magnetic resonance spectra for this enzyme. Proton resonances from histidine residues and 19F resonances from fluorine-labeled fluorotyrosine and fluorotryptophan dihydrofolate reductase have been assigned in several cases to specific amino acids in the primary sequence. Furthermore, the 31P signals from the pyrophosphate moiety of bound NADPH have been assigned and the large upfield shift for 13C-labeled (at the carboxamide carbon) NADP+ upon binding to the reductase has been explained in terms of desolvation effects.
D007780:Lacticaseibacillus casei / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D008727:Methotrexate; D008958:Models, Molecular; D009249:NADP; D011485:Protein Binding; D011487:Protein Conformation; D013762:Tetrahydrofolate Dehydrogenase; D014364:Tryptophan; D014961:X-Ray Diffraction
D009249:NADP; D014364:Tryptophan; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate