APPLICATIONS OF CALCIUM-SENSITIVE PHOTOPROTEINS IN EXPERIMENTAL BIOLOGY
APPLICATIONS OF CALCIUM-SENSITIVE PHOTOPROTEINS IN EXPERIMENTAL BIOLOGY
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DOI：
10.1111/j.1751-1097.1978.tb07624.x
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发表时间：
1978-01-01
影响因子：
3.3
通讯作者：
BLINKS, JR
中科院分区：
文献类型：
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作者：
BLINKS, JR
钙敏感性发光蛋白是“预充电的生物发光蛋白，通过结合Ca 2+或某些其他无机离子来触发发光。既不需要分子氧也不需要任何有机辅因子。第一个被描述的这种蛋白质是水母发光蛋白，由于各种原因，它是研究最广泛的蛋白质。光蛋白已被用作体外和活细胞中的Ca 2+指示剂。它们的主要优点是（1）易于信号检测，（2）高灵敏度，（3）对Ca 2+的相对特异性，和（4）缺乏毒性。发光蛋白在实验应用中的困难来自于（1）它们的一次性反应性，（2）它们的大分子尺寸，（3）它们的稀缺性，（4）实验条件对Ca 2+敏感性的影响，（5）[Ca 2 +]和光强度之间关系的非线性，以及（6）光强度跟随[Ca 2 +]突然变化的有限速度。光蛋白现在已经在二十多种类型的细胞中用作细胞内钙指示剂，并且该方法的经验正在迅速增长。它们也可用于测定体外其他物质的钙结合常数，并作为受体-配体相互作用研究的模型。
Calcium‐sensitive photoproteins are “precharged bioluminescent proteins that are triggered to emit light by binding Ca2+or certain other inorganic ions. Neither molecular oxygen nor any organic cofactor is required. The first such protein to be described was aequorin, and for various reasons that has been the one most widely studied. Photoproteins have been used as Ca2+‐indicators bothin vitroand in living cells. Their chief advantages for this are (1) ease of signal detection, (2) high sensitivity, (3) relative specificity for Ca2+, and (4) lack of toxicity. Difficulties in the experimental use of the photoproteins stem from (1) their one‐time reactivity, (2) their large molecular size, (3) their scarcity, (4) the influence of experimental conditions on the sensitivity to Ca2+, (5) the nonlinearity of the relation between [Ca2+] and light intensity, and (6) the limited speed with which light intensity follows sudden changes in [Ca2+]. Photoproteins have now been used as intracellular calcium indicators in more than two dozen types of cells, and experience with the method is rapidly growing. They are also useful in the determination of calcium binding constants for other substancesin vitro, and as models for studies of receptor‐ligand interactions in general.