Department of Biochemistry, University of Cambridge, Tennis Court Road, CB2 1QW, Cambridge, U.K..
Planta. 1977 Jan;134(2):177-81. doi: 10.1007/BF00384968.
Cellular membranes from dark grown hypocotyls of Phaseolus aureus Roxb. were separated by centrifugation on a continuous sucrose gradient. Each gradient fraction was monitored for activity of inosine diphosphatase (EC 3.6.1.6) and the ability to transfer glucose from UDP-[(14)C]glucose to endogenous lipids in vitro. The highest incorporation of radioactivity into lipids occurred in a particulate fraction correlated with the Golgi apparatus, sedimenting at sucrose densities of 31.5-33% w/w. Three endogenous lipids were glucosylated in vitro. The two main lipids were characterized as steryl glucoside and acylated steryl glucoside; data from chromatography and hydrolysis of the third lipid suggests that it is dolichyl-monophosphate-glucoside. Steryl glucoside was found to be the main glucoside synthesized, but the proportion of the acylated form increased with time. The results are discussed in the context of the role of the Golgi apparatus as a centre of membrane modification within the plant cell.
用蔗糖连续密度梯度离心法从暗培养的菜豆黄化下胚轴的细胞膜中分离出来。对每个梯度级分进行次黄嘌呤二磷酸酶(EC 3.6.1.6)活性和在体外将葡萄糖从 UDP-[(14)C]葡萄糖转移到内源性脂质的能力的监测。放射性活性掺入脂质的最高部分发生在与高尔基体相关的颗粒部分,在蔗糖密度为 31.5-33%w/w 时沉降。三种内源性脂质在体外发生葡糖基化。两种主要脂质被鉴定为甾基葡糖苷和酰化甾基葡糖苷;第三种脂质的色谱和水解数据表明它是二磷酸单磷酸葡萄糖苷。发现甾基葡糖苷是主要合成的葡糖苷,但酰化形式的比例随时间增加。结果在高尔基体作为植物细胞内膜修饰中心的作用背景下进行了讨论。