Purification and characterization of thymidylate synthetase in the liver of the mouse bearing Ehrlich ascites tumor.
The activity of thymidylate synthetase in the liver of the ddY strain male mouse increased transitorily according to the increase in tumor cell number at maximum 7-9 days after ip transplantation of Ehrlich ascites tumor. The enzyme was able to be purified from the tumor host mouse liver or from the normal mouse liver in the same manner as from tumor cells using Affi-Gel blue and methotrexate-Sepharose 4B affinity column chromatography. The three enzyme preparations obtained were purified at 27,000-38,000-, and 8,000-fold, and yielded total activities of 11, 3, and 16% of these homogenates, respectively. These preparations were similar in molecular weight to the whole enzyme (67,000) and its subunit (34,000), optimum pH, and Km values either for deoxyuridine 5'-monophosphate or tetrahydrofolate in the presence of formaldehyde. Furthermore, the amount of 5-fluoro-2'-deoxyuridine 5'-monophosphate forming the ternary complex with the enzyme and tetrahydrofolate paralleled the enzyme activities in the cytosol fractions of the three tissues. The characteristics of the tumor host liver enzyme were similar to those of the proliferating tissues, the Ehrlich ascites tumor.
Asanagi
M
M
Department of Biochemistry, Chiba University School of Medicine, Japan.
Moriyama
Y
Y
Fujimura
S
S
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D002460:Cell Line; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005468:Fluorodeoxyuridylate / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009368:Neoplasm Transplantation; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*
D005468:Fluorodeoxyuridylate; D013940:Thymidylate Synthase