Osafune T, Schiff J A, Hase E
Department of Microbiology, Tokyo Medical College, Japan.
Exp Cell Res. 1991 Apr;193(2):320-30. doi: 10.1016/0014-4827(91)90103-2.
We have localized LHCP II apoprotein in the Golgi and thylakoids of Euglena gracilis Klebs var. bacillaris Cori and strain Z Pringsheim by electron microscopy using a specific antibody and protein A-gold. Using synchronized cells (light, 14 h:dark, 10 h) we show that thylakoids are always immunoreactive. There is no reaction in the Golgi at 0 h (the beginning of the light period) but immunoreaction appears in the Golgi soon thereafter, rises to a peak at 8 h and declines to zero by 16 h (2 h into the dark period). The peak in immunoreaction in the Golgi immediately precedes the peak in cellular 14C-labeling of thylakoid LHCP II apoprotein seen by Brandt and von Kessel (Plant Physiol. (1983) 72, 616), supporting our suggestion that processing in the Golgi precedes deposition of LHCP apoprotein in the thylakoids. Substitution of preimmune serum for antiserum eliminates the immunoreaction in the Golgi, and thylakoids of synchronized cells of mutant Gr1BSL which lacks LHCP II apoprotein show no immunoreaction in the Golgi or thylakoids at any stage. Random observations indicate that the compartmentalized osmiophilic structure (COS) shows an immunoreaction with anti-LHCP II apoprotein antibody at 1 h into the light period (when the Golgi is not immunoreactive) and at 10 h into the light period (when the Golgi is fully reactive), suggesting that the COS remains immunoreactive throughout the cell cycle.
我们通过使用特异性抗体和蛋白A-金的电子显微镜技术,将光系统Ⅱ捕光叶绿素a/b结合蛋白(LHCP Ⅱ)脱辅基蛋白定位在纤细裸藻(Euglena gracilis Klebs)巴氏变种(var. bacillaris Cori)和Z品系(strain Z Pringsheim)的高尔基体和类囊体中。使用同步化细胞(光照14小时:黑暗10小时),我们发现类囊体始终具有免疫反应性。在0小时(光照期开始)高尔基体中没有反应,但此后不久高尔基体中就出现了免疫反应,在8小时达到峰值,并在16小时(黑暗期开始2小时)降至零。高尔基体中免疫反应的峰值紧接着布兰特和冯·凯塞尔(《植物生理学》(1983年)72卷,616页)所观察到的类囊体LHCP Ⅱ脱辅基蛋白细胞14C标记的峰值,这支持了我们的观点,即高尔基体中的加工过程先于LHCP脱辅基蛋白在类囊体中的沉积。用免疫前血清替代抗血清可消除高尔基体中的免疫反应,并且缺乏LHCP Ⅱ脱辅基蛋白的突变体Gr1BSL同步化细胞的类囊体在任何阶段在高尔基体或类囊体中均无免疫反应。随机观察表明,分隔嗜锇结构(COS)在光照期1小时(此时高尔基体无免疫反应)和光照期10小时(此时高尔基体完全有反应)与抗LHCP Ⅱ脱辅基蛋白抗体发生免疫反应,这表明COS在整个细胞周期中都保持免疫反应性。