Removal of Zinc from Carbonic Anhydrase. A Kinetics Experiment for Upper-Level Chemistry Laboratories
Removal of Zinc from Carbonic Anhydrase. A Kinetics Experiment for Upper-Level Chemistry Laboratories
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从碳酸酐酶中去除锌。
DOI：
10.1021/ed081p1045
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发表时间：
2004
影响因子：
3
通讯作者：
B. Adhyaru
中科院分区：
文献类型：
--
作者：
K. Williams;B. Adhyaru
碳酸酐酶活性中心的Zn（II）离子可通过与2，6-吡啶二羧酸（吡啶二羧酸）络合而除去。Zn（II）去除的动力学可以通过两种方法进行。在标准实验（适合生物物理化学，生物化学或仪器分析的学生）中，在适当的时间取出反应混合物的等分试样，并测试残留的酶活性（对硝基苯基乙酸酯水解）。在一个可选的实验（特殊项目或放射化学专业的学生）中，碳酸酐酶被标记为65 Zn。取出反应混合物的等分试样并过滤以分离释放的65 Zn，其使用液体闪烁计数器或γ计数器检测。
The Zn(II) ion in the active site of carbonic anhydrase can be removed by complexation with 2,6-pyridinedicarboxylate (dipicolinate). The kinetics of Zn(II) removal can be followed by two methods. In the standard experiment (suitable for students in biophysical chemistry, biochemistry, or instrumental analysis), aliquots of the reaction mixture are withdrawn at suitable times and tested for the residual enzymatic activity (p-nitrophenyl acetate hydrolysis). In an optional experiment (special project or for students in radiochemistry), the carbonic anhydrase is labeled with 65Zn. Aliquots of the reaction mixture are withdrawn and filtered to separate the liberated 65Zn, which is detected using either a liquid scintillation counter or a gamma counter.