Desjardins M, Morse D
Institut de Recherche en Biologie Végétale, Université de Montréal, Que., Canada.
Biochem Cell Biol. 1993 Mar-Apr;71(3-4):176-82. doi: 10.1139/o93-028.
Scintillons, the bioluminescence organelles of Gonyaulax polyedra, were purified by isopycnic density gradient centrifugation until only low levels of contaminating chloroplasts and mitochondria were detected by fluorescence and electron microscopy. Purified scintillons catalyzed the luminescent reaction with kinetics identical to those observed during the bioluminescence flash in vivo. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicated that the organelles appeared to contain only two proteins. These proteins were identified as luciferase (135 kilodaltons) and luciferin-binding protein (75 kilodaltons) based on their size and their known functions in the bioluminescence reaction in vitro. The staining of luciferin-binding protein by Coomassie blue was 2.4 +/- 0.3 (n = 19) times greater than the luciferase, suggesting that there are four binding protein monomers for every luciferase monomer. A model is proposed for the close packing of the two proteins inside the scintillons.
多甲藻（Gonyaulax polyedra）的生物发光细胞器——闪烁体，通过等密度梯度离心法进行纯化，直至通过荧光和电子显微镜仅检测到低水平的叶绿体和线粒体污染物。纯化后的闪烁体催化发光反应，其动力学与体内生物发光闪光期间观察到的动力学相同。在十二烷基硫酸钠存在下进行的聚丙烯酰胺凝胶电泳表明，这些细胞器似乎仅含有两种蛋白质。基于它们的大小及其在体外生物发光反应中的已知功能，这些蛋白质被鉴定为荧光素酶（135千道尔顿）和荧光素结合蛋白（75千道尔顿）。考马斯亮蓝对荧光素结合蛋白的染色比荧光素酶强2.4 +/- 0.3（n = 19）倍，这表明每一个荧光素酶单体对应有四个结合蛋白单体。本文提出了一个关于这两种蛋白质在闪烁体内紧密堆积的模型。
