Thymidylate synthase activity in the development of Hymenolepis diminuta.
Extracts of the tapeworm, Hymenolepis diminuta, catalyse N5,10-methylene-tetrahydrofolate-dependent release of tritium from [5-3H]dUMP, indicating the presence of thymidylate synthase. The enzyme activity was found in immature, mature and gravid proglottids, as well as in immature and mature oncospheres. The reaction showed pH optimum at 7.5. Its Michaelis constants were approximately 2 and 15 microM for dUMP and (+/-), L-N5,10-methylenetetrahydrofolate, respectively. Incubation of the tapeworm extracts with 5-F-[3H]dUMP and N5,10-methylenetetrahydrofolate resulted in formation of a labelled complex, separable under conditions of SDS polyacrylamide electrophoresis (mol. wt. of approx. 34,000), corresponding to thymidylate synthase subunit. Results of gel filtration of the above complex, under nondenaturing conditions, pointed to a dimeric structure of the enzyme.
Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warszawa.
J
J
Cieśla
Z
Z
Zieliński
B
B
Machnicka
W
W
Rode
D000818:Animals; D002850:Chromatography, Gel; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D006926:Hymenolepis / Q000201:enzymology / Q000254:growth & development*; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D013763:Tetrahydrofolates / Q000378:metabolism; D013940:Thymidylate Synthase / Q000378:metabolism*; D014316:Tritium
D003856:Deoxyuracil Nucleotides; D046911:Macromolecular Substances; D013763:Tetrahydrofolates; C013123:5,10-methylenetetrahydrofolic acid; D014316:Tritium; C007267:2'-deoxyuridylic acid; D013940:Thymidylate Synthase