Campbell A K, Dormer R L, Hallett M B
Cell Calcium. 1985 Apr;6(1-2):69-82. doi: 10.1016/0143-4160(85)90035-1.
The Ca2+-activated photoproteins aequorin and obelin are capable of detecting rapid changes in free Ca2+ over the range 10nM-100uM. Whilst they have been used to quantify free Ca transients in giant cells for some time, their use in small mammalian cells has been restricted because of the difficulty of incorporating them into live cells without impairment of cell function. We have developed three methods for incorporating photoproteins into small cells (a) reversible cell swelling (b) membrane fusion and (c) intracellular release from pinocytotic vesicles. Formation of the membrane attack complex of complement (C5b6789), via a specific cell surface antibody to activate complement, causes a rapid increase in cytoplasmic Ca2+ detectable within 5-10 s. It provides a specific method for quantifying cytoplasmic photoprotein. As a result new insights into the role of intracellular Ca2+ in cell physiology and pathology have been established.
钙离子激活的光蛋白水母发光蛋白和海仙人掌发光蛋白能够检测10纳摩尔至100微摩尔范围内游离钙离子的快速变化。虽然它们已被用于量化巨细胞中游离钙离子瞬变有一段时间了，但由于难以在不损害细胞功能的情况下将它们导入活的小哺乳动物细胞，它们在小哺乳动物细胞中的应用受到了限制。我们已经开发出三种将光蛋白导入小细胞的方法：（a）可逆性细胞肿胀；（b）膜融合；（c）从胞饮小泡中进行细胞内释放。通过一种特异性细胞表面抗体激活补体，形成补体膜攻击复合物（C5b6789），会导致在5 - 10秒内可检测到细胞质钙离子迅速增加。它提供了一种量化细胞质光蛋白的特异性方法。由此，对细胞内钙离子在细胞生理和病理中的作用有了新的认识。
