Howell K E, Palade G E
J Cell Biol. 1982 Mar;92(3):822-32. doi: 10.1083/jcb.92.3.822.
Golgi fractions isolated from rat liver homogenates have been resolved into membrane and content subfractions by treatment with 100 mM Na2CO3 pH 11.3. This procedure permitted extensive extraction of content proteins and lipoproteins, presumably because it caused an alteration of Golgi membranes that minimized the reformation of closed vesicles. The type and degree of contamination of the fractions was assessed by electron microscopy and biochemical assays. The membrane subfraction retained 15% of content proteins and lipids, and these could not be removed by various washing procedures. The content subfraction was contaminated by both membrane fragments and vesicles and accounted for 5 to 10% of the membrane enzyme activities of the original Golgi fraction. The lipid compositions of the subfractions was determined, and the phospholipids of both membrane and content were found to be uniformly labeled with [33P]phosphate administered in vivo.
从大鼠肝脏匀浆中分离出的高尔基体组分,通过用100 mM碳酸钠(pH 11.3)处理,已被分解为膜亚组分和内容物亚组分。该程序允许大量提取内容物蛋白质和脂蛋白,推测这是因为它导致高尔基体膜发生改变,从而使封闭囊泡的重新形成减到最少。通过电子显微镜和生化分析评估了各组分的污染类型和程度。膜亚组分保留了15%的内容物蛋白质和脂质,并且这些物质不能通过各种洗涤程序去除。内容物亚组分被膜碎片和囊泡污染,占原始高尔基体组分膜酶活性的5%至10%。测定了亚组分的脂质组成,发现膜和内容物的磷脂在用体内给予的[33P]磷酸盐标记后是均匀标记的。