Purification of rat liver phenylalanine hydroxylase by affinity chromatography.
Purification of rat liver phenylalanine hydroxylase by affinity chromatography.
复制标题
通过亲和层析纯化大鼠肝苯丙氨酸羟化酶。
DOI：
10.1016/0003-9861(80)90393-8
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发表时间：
1980
影响因子：
3.9
通讯作者：
Al-Janabi,JM
中科院分区：
文献类型：
--
作者：
Al-Janabi,JM
大鼠肝苯丙氨酸羟化酶已纯化到同质的全合成亲和基质。亲和基质由连接至Sepharose-4 B的琥珀酰化二氨基二丙胺臂组成，辅因子6，7-二甲基-5，6，7，8-四氢蝶呤共价连接至Sepharose-4 B。将大鼠肝匀浆的50%硫酸铵组分上柱后，用缓冲的50%乙二醇、1 mKCl一步洗脱纯酶。比活性范围为1.4至3.0单位/毫克蛋白质。已通过以下方法证明该酶是均一的：（i）不连续凝胶电泳，和（ii）十二烷基硫酸钠凝胶电泳。通过相同的技术测定亚基分子量，计算结果为51，000 - 55，000。
Rat liver phenylalanine hydroxylase has been purified to homogeneity on a totally synthesized affinity matrix. The affinity matrix consisted of a succinylated diaminodipropylamine arm linked to Sepharose-4B, to which the cofactor, 6,7-dimethyl-5,6,7,8-tetrahydropterin, was covalently linked. The pure enzyme was eluted with buffered 50% ethylene glycol, 1mKCl in one step after the 50% ammonium sulfate fraction of the rat liver homogenate was applied to the affinity column. Specific activities ranging from 1.4 to 3.0 units/mg of protein were obtained. The enzyme has been shown to be homogeneous by: (i) discontinuous gel electrophoresis, and (ii) sodium dodecyl sulfate gel electrophoresis. The subunit molecular weight was determined by the same technique and was calculated to be between 51,000 and 55,000.