Purification of thymidylate synthetase from Ehrlich ascites carcinoma cells.
作者： Arnold Fridland , Robert J. Langenbach , Charles Heidelberger
DOI: 10.1016/S0021-9258(19)45860-2
关键词: Ehrlich ascites carcinoma 、 Sephadex 、 Size-exclusion chromatography 、 Biochemistry 、 Metabolism 、 Enzyme 、 Thymidylate synthase 、 Biology 、 Sodium dodecyl sulfate 、 Polyacrylamide gel electrophoresis
摘要: Abstract Thymidylate synthetase has been purified about 4,700-fold from Ehrlich ascites carcinoma cells. The procedures used to isolate the enzyme were ammonium sulfate fractionation, DEAE-cellulose chromatography, Sephadex G-100 filtration, and chromatography on Brushite. was demonstrated be homogeneous by disc electrophoresis polyacrylamide gel. By use of gel filtration sodium dodecyl electrophoresis, we estimated molecular weight protein 67,000 71,000 respectively. did not show a requirement for Mg2+. Km value dl, l-methylenetetrahydrofolate 43 µm deoxyuridylate 6.3 µm. Thiols enhanced activity at all stages purification.
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