Physico-chemical properties of thymidylate synthase from Lactobacillus leichmannii: thiol groups, amino acid composition and the terminal end-groups.

ABSTRACT

Thymidylate synthase (5,10-methylenetetrahydrofolate deoxyuridylate C-methyltransferase, EC 2.1.1.45) from Lactobacillus leichmannii was completely inactivated after 5 min of heat treatment at 55 degrees C. A remarkable synergistic effect with no loss in activity was noted when 10(-3) M dUMP was added to the enzyme before subjecting to heat treatment. The enzyme got activated in the presence of 2-mercaptoethanol (75 mM) and inhibited by pCMB (I50 = 5 microM). It had 2 free sulfhydryl groups and a single disulfide bond. The two identical subunits of the 74 kDa dimer were possibly bonded by a single disulfide linkage. It had a total of 652 amino acids with methionine as the amino-terminal and alanine as the carboxy-terminal amino acid residues. The carboxy-terminal end-group alanine was preceded by valine, lysine and proline sequentially in that order.