阿拉伯糖基：羟脯氨酸转移酶在马铃薯组织培养细胞内膜系统中的定位。
The location of arabinosyl:hydroxyproline transferase in the membrane system of potato tissue culture cells.
作者信息
Owens R J, Northcote D H
出版信息
Biochem J. 1981 Jun 1;195(3):661-7. doi: 10.1042/bj1950661.
Abstract
Incubation of a particulate preparation from potato tissue culture cells with UDP-beta-L-[1-3H] arabinose yielded a glycoprotein fraction containing labelled material with the characteristics of hydroxyproline arabinosides. The sugar-protein linkage was resistant to hot alkaline hydrolysis, and the hydrolytic products showed similar electrophoretic and chromatographic behavior to authentic hydroxyproline-arabinosides prepared from potato tissue culture cell walls. Incorporation of arabinose into glycoprotein was stimulated by the addition of de-arabinosylated potato lectin. The product of the incubation co-migrated with native potato lectin on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The subcellular distribution of the arabinosyl-transferase was investigated by fractionating potato tissue culture membranes on a discontinuous sucrose gradient in the presence or absence of Mg2+. Under both fractionation conditions the highest specific activity of the enzyme was found in the Golgi-enriched fraction. The results are discussed in relation to the synthesis of the hydroxy-proline-rich glycoprotein component of plant cell walls.
摘要
用UDP-β-L-[1-³H]阿拉伯糖培养马铃薯组织培养细胞的颗粒制剂，得到了一个糖蛋白组分，该组分含有具有羟脯氨酸阿拉伯糖苷特征的标记物质。糖-蛋白连接键对热碱水解具有抗性，水解产物在电泳和色谱行为上与从马铃薯组织培养细胞壁制备的 authentic 羟脯氨酸-阿拉伯糖苷相似。添加去阿拉伯糖基化的马铃薯凝集素可刺激阿拉伯糖掺入糖蛋白。孵育产物在十二烷基硫酸钠/聚丙烯酰胺凝胶电泳上与天然马铃薯凝集素共迁移。通过在存在或不存在Mg²⁺的情况下在不连续蔗糖梯度上分离马铃薯组织培养膜，研究了阿拉伯糖基转移酶的亚细胞分布。在两种分级分离条件下，在富含高尔基体的组分中发现该酶的比活性最高。结合植物细胞壁中富含羟脯氨酸的糖蛋白成分的合成对结果进行了讨论。
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