Variant of Perivitelline Membrane Glycoprotein ZPC of Japanese Quail (Coturnix japonica) Lacking Its Cytoplasmic Tail Exhibits the Retention in the Endoplasmic Reticulum of Chinese Hamster Ovary (CHO-K1) Cells1
Variant of Perivitelline Membrane Glycoprotein ZPC of Japanese Quail (Coturnix japonica) Lacking Its Cytoplasmic Tail Exhibits the Retention in the Endoplasmic Reticulum of Chinese Hamster Ovary (CHO-K1) Cells1
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缺少胞质尾的日本鹌鹑 (Coturnix japonica) 卵周膜糖蛋白 ZPC 变体在中国仓鼠卵巢 (CHO-K1) 细胞内质网中表现出滞留作用1
DOI：
10.1095/biolreprod.103.018333
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发表时间：
2003
期刊：
影响因子：
3.5
通讯作者：
M. Mori
中科院分区：
文献类型：
--
作者：
T. Sasanami;A. Hanafy;M. Toriyama;M. Mori
摘要鸟类卵黄周膜是一种与哺乳动物卵透明膜同源的膜，由至少两种糖蛋白组成。我们以前的研究表明，它的成分之一，ZPC，这是在卵巢颗粒细胞合成的，被分泌后的羧基末端蛋白水解加工，这一事件是一个先决条件ZPC分泌鹌鹑。在本研究中，我们研究了细胞质尾，这是成功地删除后，蛋白水解处理，在膜运输，蛋白水解处理，和鹌鹑ZPC的分泌的作用。在追求这一点，我们产生了一个截短的ZPC突变体缺乏位于其C-末端的胞质尾，并检查其在哺乳动物细胞系中的表达。蛋白质印迹分析表明，细胞质尾部缺陷的ZPC既不分泌，也不经历细胞中的蛋白水解加工。免疫荧光分析和收购的细胞质尾缺陷ZPC的内切糖苷酶H消化的阻力表明，删除的细胞质尾干扰的蛋白质从内质网到高尔基体的细胞内运输。这些结果表明，鹌鹑ZPC的胞质尾区可能具有负责新合成的ZPC从内质网到高尔基体的有效运输的决定因素。
Abstract Avian perivitelline membrane, an investment homologous to the mammalian zona pellucida, is composed of at least two glycoproteins. Our previous studies demonstrated that one of its components, ZPC, which is synthesized in the ovarian granulosa cells, is secreted after carboxy-terminal proteolytic processing, and this event is a prerequisite event for ZPC secretion in quail. In the present study, we examined the role of the cytoplasmic tail, which is successfully removed after proteolytic processing, in membrane transport, proteolytic processing, and the secretion of quail ZPC. In pursuit of this, we produced a truncated ZPC mutant lacking the cytoplasmic tail located in its C-terminus and examined its expression in the mammalian cell line. Western blot analyses demonstrated that the cytoplasmic tail-deficient ZPC was neither secreted nor underwent proteolytic processing in the cells. Immunofluorescence analysis and the acquisition of resistance to endoglycosidase H digestion of the cytoplasmic tail-deficient ZPC demonstrated that the deletion of the cytoplasmic tail interferes with the intracellular trafficking of the protein from the endoplasmic reticulum to the Golgi apparatus. These results indicate that the cytoplasmic tail of quail ZPC might possess the determinant responsible for the efficient transport of the newly synthesized ZPC from the endoplasmic reticulum to the Golgi apparatus.
DOI：
10.1016/s0021-9258(18)61070-1
发表时间：
1987-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
P. Matsudaira
通讯作者：
P. Matsudaira
影响因子：
3.3
作者：
H. Qi;Zev Williams;P. M. Wassarman
通讯作者：
H. Qi;Zev Williams;P. M. Wassarman