来自培养哺乳动物细胞的二氢叶酸还原酶的纯化及性质
Purification and properties of dihydrofolate reductase from cultured mammalian cells.
作者信息
Gauldie J, Marshall L, Hillcoat B L
出版信息
Biochem J. 1973 Jun;133(2):349-56. doi: 10.1042/bj1330349.
Abstract
Dihydrofolate reductase was purified quickly and simply from small quantities of cultured mammalian cells by affinity chromatography. On gel electrophoresis of the purified enzyme, multiple bands of activity resulted from enzyme-buffer interaction at low but not high buffer concentration. A Ferguson plot (Ferguson, 1964) showed that this heterogeneity was due to a charge difference with no alteration in the size of the enzyme. Stimulation of enzyme activity by KCl, urea and p-hydroxymercuribenzoate, and inhibition by methotrexate and trimethoprim, showed only minor differences between the various enzymes.
摘要
通过亲和层析法可从小量培养的哺乳动物细胞中快速简便地纯化二氢叶酸还原酶。对纯化酶进行凝胶电泳时，在低缓冲液浓度而非高缓冲液浓度下，酶与缓冲液的相互作用产生了多条活性带。弗格森作图法（弗格森，1964年）表明，这种异质性是由于电荷差异导致的，酶的大小没有改变。氯化钾、尿素和对羟基汞苯甲酸对酶活性有刺激作用，而甲氨蝶呤和甲氧苄啶则有抑制作用，不同酶之间仅表现出微小差异。
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