Lysosomal membrane dynamics: structure and interorganellar movement of a major lysosomal membrane glycoprotein.
Lysosomal membrane dynamics: structure and interorganellar movement of a major lysosomal membrane glycoprotein.
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DOI：
10.1083/jcb.102.5.1593
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发表时间：
1986-05
期刊：
影响因子：
--
通讯作者：
Fambrough DM
中科院分区：
文献类型：
--
作者：
Lippincott-Schwartz J;Fambrough DM
用单克隆抗体技术研究了鸡成纤维细胞溶酶体中一种完整膜糖蛋白的生物化学和胞内转运。糖蛋白的表观分子量为95000 ~ 105000。结构分析包括[35S]蛋氨酸代谢标记和糖苷酶裂解，发现存在许多寡糖链n -连接到一个表观分子量为48,000的核心多肽。抗体结合位点与吖啶橙吸收区重合，溶酶体样液泡膜内表面的电子免疫细胞化学标记，以及糖蛋白与Percoll梯度中富含溶酶体的亚细胞组分优先结合，证明了糖蛋白在溶酶体上的初步定位。此外，在内吞囊泡和质膜上检测到少量的糖蛋白。为了研究糖蛋白的细胞内通路，我们使用了一种单克隆抗体，该抗体与糖蛋白在细胞表面的结合对抗原分子的数量和亚细胞分布没有影响。单克隆抗体在37℃下孵育鸡成纤维细胞，导致抗体迅速被吸收并随后被递送到溶酶体，在溶酶体中抗体被降解。这一过程在连续暴露于抗体的细胞上持续数小时不减弱，并且不被添加环己亚胺阻断。预先标记抗体的细胞的质膜中抗原位点的补充速率（t1/2 = 2 min）基本上等于抗体结合到细胞表面的内在化速率。这些结果表明，这种糖蛋白在质膜和内体和/或溶酶体的膜之间有一个连续和快速的交换。
The biochemistry and intracellular transit of an integral membrane glycoprotein of chicken fibroblast lysosomes were studied with monoclonal antibody techniques. The glycoprotein had an apparent molecular weight of 95,000-105,000. Structural analysis involving metabolic labeling with [35S]methionine and cleavage with glycosidases revealed the presence of numerous oligosaccharide chains N-linked to a core polypeptide of apparent molecular weight 48,000. A primary localization of the glycoprotein to lysosomes was demonstrated by the coincidence of antibody binding sites with regions of acridine orange uptake, electron immunocytochemical labeling on the inner surface of lysosome-like vacuolar membranes, and preferential association of the glycoprotein with lysosome-enriched subcellular fractions from Percoll gradients. In addition, small quantities of the glycoprotein were detected on endocytic vesicle and plasma membranes. To study the intracellular pathway of the glycoprotein, we used a monoclonal antibody whose binding to the glycoprotein at the cell surface had no effect on the number or subcellular distribution of antigen molecules. Incubation of chicken fibroblasts with monoclonal antibody at 37 degrees C led to the rapid uptake and subsequent delivery of antibody to lysosomes, where antibody was degraded. This process continued undiminished for many hours on cells continuously exposed to the antibody and was not blocked by the addition of cycloheximide. The rate at which antigen sites were replenished in the plasma membrane of cells prelabeled with antibody (t1/2 = 2 min) was essentially equivalent to the rate of internalization of antibody bound to cell surfaces. These results suggest that there is a continuous and rapid exchange of this glycoprotein between plasma membrane and the membranes of endosomes and/or lysosomes.
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溶酶体膜的糖蛋白。
影响因子：
7.8
作者：
LEWIS, V;GREEN, SA;MARSH, M;VIHKO, P;HELENIUS, A;MELLMAN, I
通讯作者：
MELLMAN, I
影响因子：
7.8
作者：
CHEN, JW;MURPHY, TL;WILLINGHAM, MC;PASTAN, I;AUGUST, JT
通讯作者：
AUGUST, JT
DOI：
10.1073/pnas.75.7.3327
发表时间：
1978-01-01
影响因子：
11.1
作者：
OHKUMA, S;POOLE, B
通讯作者：
POOLE, B
影响因子：
2.4
作者：
KARTNER, N;ALON, N;RIORDAN, JR
通讯作者：
RIORDAN, JR
影响因子：
64.5
作者：
ROME, LH;GARVIN, AJ;NEUFELD, EF
通讯作者：
NEUFELD, EF