Shimomura O, Musicki B, Kishi Y, Inouye S
Marine Biological Laboratory, Woods Hole, Massachusetts.
Cell Calcium. 1993 May;14(5):373-8. doi: 10.1016/0143-4160(93)90041-4.
15 kinds of recombinant semi-synthetic aequorins and a recombinant fluorescein-conjugated aequorin were prepared and their properties in Ca(2+)-triggered luminescence were studied. The semi-synthetic aequorins showed a wide range of Ca(2+)-sensitivity. The luminescence intensity of a high-sensitivity type (hcp-aequorin) was greater than 10(4)-times that of a low-sensitivity type (n-aequorin) at pCa 6.0-6.5. The fluorescein-conjugated aequorin exhibited fluorescence in addition to the Ca(2+)-triggered luminescence, thus it can be used to visualize the diffusion and distribution of aequorin in cells. The data obtained, particularly the Ca(2+)-sensitivity curves, are useful in selecting a suitable semi-synthetic aequorin for an experiment.
制备了15种重组半合成水母发光蛋白和1种重组荧光素偶联水母发光蛋白,并研究了它们在Ca(2+)触发发光中的特性。半合成水母发光蛋白表现出广泛的Ca(2+)敏感性。在pCa 6.0 - 6.5时,高敏感性类型(hcp-水母发光蛋白)的发光强度比低敏感性类型(n-水母发光蛋白)高10(4)倍以上。荧光素偶联水母发光蛋白除了Ca(2+)触发发光外还表现出荧光,因此可用于可视化水母发光蛋白在细胞中的扩散和分布。所获得的数据,特别是Ca(2+)敏感性曲线,对于选择适合实验的半合成水母发光蛋白很有用。