Rapid purification of mitochondrial hexokinase from rat brain by a single affinity chromatography step on Affi-Gel blue.
Rapid purification of mitochondrial hexokinase from rat brain by a single affinity chromatography step on Affi-Gel blue.
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通过 Affi-Gel blue 上的单一亲和层析步骤快速纯化大鼠脑中的线粒体己糖激酶。
DOI：
10.1080/10826068908544893
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Wilson,JE
中科院分区：
文献类型：
--
作者：
Wilson,JE
大鼠脑线粒体己糖激酶是在葡萄糖6-磷酸作用下选择性地从线粒体中释放出来的，通过Affi-Gel Blue单步亲和层析纯化，均匀性大于90%；与该基质结合的Cibacron Blue F3GA配体充当ATP的类似物，ATP是酶的正常底物，使用葡萄糖6-磷酸完成选择性洗脱，葡萄糖6-磷酸是与ATP竞争的配体。通过对先前描述的过程的这种和其他修改（参考文献8），可以很容易地获得高纯度的酶，产量很高，并且保留了与线粒体重新结合的能力。
The mitochondrial hexokinase from rat brain, selectively released from mitochondria by the action of glucose 6-phosphate, can be purified to greater than 90% homogeneity by a single affinity chromatography step on Affi-Gel Blue; the Cibacron Blue F3GA ligand bound to this matrix serves as an analog of ATP, the normal substrate for the enzyme, and selective elution is accomplished using glucose 6-phosphate which is a competitive ligand vs. ATP. With this and other modifications to the previously described procedure (Ref. 8), highly purified enzyme is readily obtained in good yield and with retention of the ability to rebind to mitochondria.