Brada D, Schekman R
Department of Biochemistry, University of California, Berkeley 94720.
J Bacteriol. 1988 Jun;170(6):2775-83. doi: 10.1128/jb.170.6.2775-2783.1988.
Immunoelectron microscopy of Saccharomyces cerevisiae cells embedded in Lowicryl K4M has been used to localize invertase and plasma membrane (PM) ATPase in secretory organelles. sec mutant cells incubated at 37 degrees C were prepared for electron microscopy, and thin sections were incubated with polyclonal antibodies, followed by decoration with protein A-gold. Specific labeling of invertase was seen in the lumen of the endoplasmic reticulum, Golgi apparatus, and secretory vesicles in mutant cells that exaggerate these organelles. PM ATPase accumulated within the same organelles. Double-immune labeling revealed that invertase and PM ATPase colocalized in secretory vesicles. These results strengthen the view that secretion and plasma membrane assembly are biosynthetically coupled in yeast.
对包埋在低温包埋剂K4M中的酿酒酵母细胞进行免疫电子显微镜观察,以将转化酶和质膜(PM)ATP酶定位在分泌细胞器中。将在37摄氏度下孵育的sec突变细胞制备用于电子显微镜观察,薄切片用多克隆抗体孵育,然后用蛋白A-金进行标记。在夸大这些细胞器的突变细胞的内质网、高尔基体和分泌小泡腔中可见转化酶的特异性标记。PM ATP酶在相同的细胞器中积累。双重免疫标记显示转化酶和PM ATP酶共定位于分泌小泡中。这些结果强化了这样一种观点,即酵母中的分泌和质膜组装在生物合成上是偶联的。