Brett C T, Northcote D H
Biochem J. 1975 Apr;148(1):107-17. doi: 10.1042/bj1480107.
Membrane fractions were obtained from peas roots by using a method that permitted the isolation of a fraction rich in relatively intact dictyosome stacks. No chemical fixatives were used. The method involved incubation of the roots with cellulase, followed by gentle homogenization and sucrose-density-gradient fractionation of the homogenate. The fractions were characterized by electron microscopy. All fractions were enzymically active in incorporating glucose from UDP-glucose into water-insoluble glycolipids containing both single glucose residues and glucose oligosaccharides. Some or all of the linkages of glucose to lipid were through phosphate esters. A substance containing glucose oligosaccharides attached to or very strongly adsorbed on to protein was also formed. The membrane fractions also incorporated glucose from UDP-glucose into alkali-soluble and alkali-insoluble beta-glucans, which like the oligosaccharides contained beta(1leads to 3) and beta-(1leads to4) linkages. The distribution of the enzymic activities and the chemical properties of the lipid-linked and protein-linked oligosaccharides suggest that they may be intermediates in beta-glucan synthesis. The synthetic activity is associated with smooth-membrane vesicles which may be derived from the plasma membrane.
通过一种方法从豌豆根中获得膜组分，该方法能够分离出富含相对完整的高尔基体堆叠的组分。未使用化学固定剂。该方法包括将根与纤维素酶一起孵育，随后进行温和匀浆并对匀浆进行蔗糖密度梯度分级分离。通过电子显微镜对各组分进行表征。所有组分在将UDP - 葡萄糖中的葡萄糖掺入含有单个葡萄糖残基和葡萄糖寡糖的水不溶性糖脂方面均具有酶活性。葡萄糖与脂质的部分或所有连接是通过磷酸酯。还形成了一种含有附着在蛋白质上或非常强烈吸附在蛋白质上的葡萄糖寡糖的物质。膜组分还将UDP - 葡萄糖中的葡萄糖掺入碱溶性和碱不溶性β - 葡聚糖中，这些葡聚糖与寡糖一样含有β(1→3)和β-(1→4)连接。酶活性的分布以及脂质连接和蛋白质连接的寡糖的化学性质表明它们可能是β - 葡聚糖合成的中间体。合成活性与可能源自质膜的光滑膜泡相关。
