On the mechanism of 2'-deoxyuridylate hydroxymethylase.
dUMP hydroxymethylase from SP01-infected Bacillus subtilis has been purified 160-fold by chromatography on DEAE-cellulose and ethylagarose. The enzyme catalyzes exchange of the 5-hydrogen of dUMP for protons of water in the presence or absence of the cofactor CH2-H4folate. Upon treatment with FdUMP and CH2-H4folate, an isolable covalent complex is formed which is believed to be structurally similar to a steady-state intermediate of the normal reaction. The FdUMP-CH2-H4folate-dUMP hydroxymethylase complex is stable toward denaturation with sodium dodecyl sulfate and shows a subunit molecular weight of 46 000. By analogy with chemical models and studies of dTMP synthetase, a mechanism is proposed for the reaction catalyzed by dUMP hydroxymethylase.
D001412:Bacillus subtilis / Q000201:enzymology*; D002384:Catalysis; D055598:Chemical Phenomena; D002621:Chemistry; D002848:Chromatography, DEAE-Cellulose; D019877:Hydroxymethyl and Formyl Transferases; D013940:Thymidylate Synthase / Q000378:metabolism; D014166:Transferases / Q000302:isolation & purification / Q000378:metabolism*
D014166:Transferases; D013940:Thymidylate Synthase; D019877:Hydroxymethyl and Formyl Transferases; C023116:dUMP-hydroxymethylase