Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation.
Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation.
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天冬酰胺连接的寡糖加工和溶酶体酶磷酸化的广泛亚细胞组织的证据。
DOI：
10.1016/s0021-9258(18)32843-6
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
S. Kornfeld
中科院分区：
文献类型：
--
作者：
D. Goldberg;S. Kornfeld
从小鼠淋巴瘤BW5147制备的膜。3细胞和P388D1巨噬细胞在连续的蔗糖梯度上分级，并检测与天冬酰胺连接的寡糖加工有关的酶。这些酶从致密膜到轻膜的分布顺序与糖蛋白生物合成中既定的事件顺序相关。一些以前定位于高尔基体的酶在梯度上分成四个区域。UDP-N-乙酰氨基葡萄糖：溶酶体酶N-乙酰氨基葡萄糖-磷酸转移酶是一种催化溶酶体酶的高甘露低聚糖选择性磷酸化的酶，存在于最致密的膜中。其次是N-乙酰氨基葡萄糖L-磷酸二酯-AN-乙酰氨基葡萄糖苷酶。与天冬酰胺连接低聚糖加工后期相关的几种酶定位于第三区。UDP-半乳糖：N-乙酰谷氨酸半乳糖基转移酶存在于最轻的膜上(区域IV)。利用[2-~3H]甘露糖的脉冲追逐实验表明，标记的天冬酰胺连接的寡糖中间体在体内的分布与这些加工酶的分布有关。对溶酶体酶的磷酸化低聚糖的分析表明，这些酶已经通过了含有半乳糖基转移酶和唾液酸基转移酶的高尔基体区域。
Membranes prepared from mouse lymphoma BW5147. 3 cells and P388D1 macrophages were fractionated on a continuous sucrose gradient and assayed for enzymes involved in the processing of asparaginelinked oligosaccharides. The order in which these enzymes distributed from dense to light membranes correlated with the established sequence of events in glycoprotein biosynthesis. A number of enzymes which have been previously localized to the Golgi separated into four regions on the gradient. UDP-N-acetylglucosamine: lysosomal enzyme N-acetylglucosamine-lphosphotransferase, the enzyme which catalyzes the selective phosphorylation of the high mannose oligosaccharides of lysosomal enzymes, was present in the densest membranes. N-Acetylglucosamine l-phosphodiester aN-acetylglucosaminidase was in the next region. Several enzymes involved in the late stages of asparagine-linked oligosaccharide processing were localized to the third region. UDP-ga1actose: N-acetylglu-cosamine galactosyltransferase was present in the lightest membranes (region IV). Pulse-chase experiments utilizing [2-3H] mannose demonstrated that the distribution of in vivo labeled asparagine-linked oligosaccharide intermediates correlates with the distribution of these processing enzymes. Analysis of the phosphorylated oligosaccharides of lysosomal enzymes which were bound to the phosphomannosyl receptor indicated that these enzymes had already passed through the region of the Golgi which contains galactosyltransferase and sialyltransferase.