Isolation of the covalent binary complex of 5-fluorodeoxyuridylate and thymidylate synthetase by trichloroacetic acid precipitation.
Isolation of the covalent binary complex of 5-fluorodeoxyuridylate and thymidylate synthetase by trichloroacetic acid precipitation.
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通过三氯乙酸沉淀分离 5-氟脱氧尿苷酸和胸苷酸合成酶的共价二元复合物。
DOI：
10.1016/0006-291x(84)90912-4
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发表时间：
1984
影响因子：
3.1
通讯作者：
Dunlap,RB
中科院分区：
文献类型：
--
作者：
Moore,MA;Ahmed,F;Dunlap,RB
利用三氯乙酸沉淀法分离出5-氟脱氧尿苷酸和胸苷酸合成酶之间的共价二元配合物，为该配合物的存在提供了强有力的化学证据。这一结果与n -乙基马来酰亚胺灭活胸腺酸合成酶的对照实验结果表明，只有与蛋白质共价结合的核苷酸在沉淀物的反复洗涤中存活下来。在此条件下，共价二元配合物的最大结合化学计量量约为0.9,Kd= 1.1 × 10−5M。此外，亚甲基四氢叶酸-5-氟脱氧尿苷-胸苷酸合成酶三元配合物的结合率为1.7。
Strong chemical evidence for the existence of a covalent binary complex between 5-fluorodeoxyuridylate and thymidylate synthetase was provided by the isolation of the complex by trichloroacetic acid precipitation. This result together with that of a control experiment with N-ethymaleimide inactivated thymidylate synthetase demonstrated that only nucleotide covalently bound to the protein survived repeated washings of the precipitate. Under the conditions used, a maximum binding stoichiometry of about 0.9 was obtained for the covalent binary complex, Kd= 1.1 × 10−5M. Also, a binding ratio of 1.7 was obtained for the methylenetetrahydrofolate-5-fluorodeoxyuridylate-thymidylate synthetase ternary complex.