Purification of guanosine triphosphate cyclohydrolase I and dihydrofolate reductase on a dihydrofolate-Sepharose affinity column.
Purification of guanosine triphosphate cyclohydrolase I and dihydrofolate reductase on a dihydrofolate-Sepharose affinity column.
复制标题
在二氢叶酸-琼脂糖亲和柱上纯化三磷酸鸟苷环水解酶 I 和二氢叶酸还原酶。
DOI：
10.1016/0003-2697(79)90120-9
复制
发表时间：
1979
影响因子：
2.9
通讯作者：
R. Then
中科院分区：
文献类型：
--
作者：
R. Then
Folate was coupled to AH-Sepharose 4B, the gel poured into small columns, and the Sepharose-bound folate reduced in situ to dihydrofolate by dithionite/ascorbate at pH 6 to 7. The dihydrofolate-Sepharose column was used to purify guanosine triphosphate cyclohydrolase I (EC 3.5.4.16) and dihydrofolate reductase (EC 1.5.1.3). All steps were carried out in the cold and in the presence of 20 mm mercaptoethanol. GTP cyclohydrolase I bound strongly to the dihydrofolate-Sepharose column and was purified several-hundred-fold in a single step. It did not bind to folate-Sepharose. Binding to dihydrofolate-Sepharose is assumed to reflect a physiological role of dihydrofolate. GTP cyclohydrolase II did not bind to either folate- or dihydrofolate-Sepharose. Dihydrofolate reductase from Escherichia coli B and from rat liver did not bind to folate-Sepharose under the test conditions, but could be well purified on the dihydrofolate-Sepharose column. This column is judged to beuseful for the purification of other folate-converting enzymes.
