15N and 1H NMR evidence for multiple conformations of the complex of dihydrofolate reductase with its substrate, folate.
The binding of folate to Lactobacillus casei dihydrofolate reductase in the presence and absence of NADP+ has been studied by 15N NMR, using [5-15N]folate. In the presence of NADP+, three separate signals were observed for the single 15N atom, in agreement with our earlier evidence from 1H and 13C NMR for multiple conformations of this complex [(1982) Biochemistry 21, 5831-5838]. The 15N spectra of the binary enzyme-folate complex provide evidence for the first time that this complex also exists in at least two conformational states. This is confirmed by the observation of two separate resonances for the 7-proton of bound folate, located by two-dimensional exchange spectroscopy.
D001426:Bacterial Proteins / Q000378:metabolism*; D005492:Folic Acid / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007780:Lacticaseibacillus casei / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D009249:NADP / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism*
D001426:Bacterial Proteins; D009249:NADP; D005492:Folic Acid; D013762:Tetrahydrofolate Dehydrogenase