Site-specific mutagenesis of the calcium-binding photoprotein aequorin.
Site-specific mutagenesis of the calcium-binding photoprotein aequorin.
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钙结合发光蛋白水母发光蛋白的位点特异性诱变。
DOI：
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发表时间：
1986
影响因子：
11.1
通讯作者：
Y. Sakaki
中科院分区：
文献类型：
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作者：
F. I. Tsuji;S. Inouye;T. Goto;Y. Sakaki
来自水母Aequoria维多利亚的发光蛋白水母发光蛋白在微量Ca（2+）存在下通过分子内反应发光。为了理解反应的机制，进行了结构-功能关系的研究，以修饰其某些氨基酸残基。这是通过进行脱辅基水母发光蛋白cDNA的阿托西肽定向位点特异性诱变并在大肠杆菌中表达诱变的cDNA来完成的。在三个Ca（2+）结合位点、三个半胱氨酸和一个疏水区中的组氨酸处进行氨基酸取代。随后对修饰的水母发光蛋白的分析表明，Ca（2+）结合位点、半胱氨酸和可能的组氨酸都在水母发光蛋白的生物发光反应中起作用。
The luminescent protein aequorin from the jellyfish Aequoria victoria emits light by an intramolecular reaction in the presence of a trace amount of Ca(2+). In order to understand the mechanism of the reaction, a study of structure-function relationships was undertaken with respect to modifying certain of its amino acid residues. This was done by carrying out oligonucleotide-directed site-specific mutagenesis of apoaequorin cDNA and expressing the mutagenized cDNA in Escherichia coli. Amino acid substitutions were made at the three Ca(2+)-binding sites, the three cysteines, and a histidine in one of the hydrophobic regions. Subsequent assay of the modified aequorin showed that the Ca(2+)-binding sites, the cysteines, and probably the histidine all play a role in the bioluminescence reaction of aequorin.