Study of thymidylate synthetase-function by laser Raman spectroscopy.
The Laser-Raman spectra of thymidylate synthetase have been obtained with 488 nm excitation from an argon ion laser. Raman bands observed in the range 600-800 cm-minus-1 have been assigned to functional groups of constituent amino acids. The band positions and intensities in the Amide I (1600-1700 cm-minus-1) and Amide III (1200-1300 cm-minus-1) regions, suggest that the enzyme is a mixture of alpha-helical and unordered conformations. Low levels of beta-structure cannot be excluded. The spectra of the ternary complex formed by reacting thymidylate synthetase with (+)-L-methylenetetrahydrofolate and fluorodeoxyuridylate reveals a new band at 1618 cm-minus-1 assigned to the C=N stretching vibration. This band may be due to formation of dihydrofolate or an iminium ion. The overall secondary structure of thymidylate synthetase does not change on formation of the ternary complex. However, the spectrum of the complex indicates local changes in groups such as ionized carboxyl (1400 cm-minus-1), tryptophan (1003 cm-minus-1)  and CH-3, CH-2 deformation modes (1440-1470 cm-minus-1).
D001665:Binding Sites; D004352:Drug Resistance, Microbial; D005468:Fluorodeoxyuridylate; D007780:Lacticaseibacillus casei / Q000187:drug effects / Q000201:enzymology; D007834:Lasers; D008727:Methotrexate / Q000494:pharmacology; D008780:Methyltransferases; D011485:Protein Binding; D011487:Protein Conformation; D012542:Scattering, Radiation; D013055:Spectrophotometry, Infrared; D013057:Spectrum Analysis; D013763:Tetrahydrofolates; D013940:Thymidylate Synthase / Q000378:metabolism
D013763:Tetrahydrofolates; D005468:Fluorodeoxyuridylate; D008780:Methyltransferases; D013940:Thymidylate Synthase; D008727:Methotrexate