ON ROLE OF PYRIDOXAL 5'-PHOSPHATE IN PHOSPHORYLASE .2. RESOLUTION OF RABBIT MUSCLE PHOSPHORYLASE B
ON ROLE OF PYRIDOXAL 5'-PHOSPHATE IN PHOSPHORYLASE .2. RESOLUTION OF RABBIT MUSCLE PHOSPHORYLASE B
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DOI：
10.1021/bi00870a044
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发表时间：
1966-01-01
期刊：
影响因子：
2.9
通讯作者：
FISCHER, EH
中科院分区：
文献类型：
--
作者：
SHALTIEL, S;HEDRICK, JL;FISCHER, EH
本文报道了一种从兔肌磷酸化酶B中除去吡哆醛5[image]-磷酸（PLP）的方法。该程序涉及2个同时操作，即，通过暴露共价结合的PLP的适当变形剂使蛋白质变形，以及通过与PLP试剂相互作用除去辅因子。测试了许多变形缓冲剂，其中咪唑鎓柠檬酸盐被证明是最有效的。使用该缓冲液和L-半胱氨酸作为羰基试剂，在pH 6.2，0[度]，
A method for the removal of pyridoxal 5[image]-phosphate (PLP) from rabbit muscle phosphorylase b is described. The procedure involves 2 simultaneous operations, namely, distortion of the protein by an appropriate deforming agent which exposes the covalently bound PLP, and removal of thecofactor by interaction with a PLP reagent. A number of deforming buffers were tested, of which imidazolium citrate proved to be the most effective. Using this buffer and L-cysteine as the carbonyl reagent, resolution of phosphorylase occurred with a half-life of about 5 min. at pH 6.2, 0[degree],