Endoplasmic reticulum retention and prolonged association of a von Willebrand's disease-causing von Willebrand factor variant with ERp57 and calnexin
Endoplasmic reticulum retention and prolonged association of a von Willebrand's disease-causing von Willebrand factor variant with ERp57 and calnexin
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DOI：
10.1006/bbrc.2000.4139
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发表时间：
2001-01-19
影响因子：
3.1
通讯作者：
Daly, ME
中科院分区：
文献类型：
--
作者：
Allen, S;Goodeve, AC;Daly, ME
我们以前已经确定了一个突变（R273 W）的血管性血友病因子（VWF）前肽，导致定量缺乏血浆VWF和损失的高分子量VWF多聚体。在COS-7细胞中表达的具有R273 W突变的重组VWF（rVWFR 273 W）在细胞内位置显示出严重受损的分泌和降解[艾伦，S.，等人（2000）Blood 96，560-568]。在这份报告中，我们使用脉冲追踪分析和内切糖苷酶H消化的野生型rVWF和rVWFR 273 W免疫沉淀的COS-7细胞显示，rVWFR 273 W保留在内质网（ER）。我们首次证明野生型rVWF和rVWFR 273 W在ER脉冲追踪分析中的生物合成期间与巯基依赖性氧化还原酶ERp 57相互作用，表明与野生型rVWF相比，rVWFR 273 W与ERp 57和钙连接蛋白的相互作用延长。相反，rVWFR 273 W和野生型rVWF在与钙网蛋白相互作用的时间过程中没有明显差异。(C)北京：科学出版社.
We have previously identified a mutation (R273W) in the von Willebrand factor (VWF) propeptide that results in quantitative deficiency of plasma VWF and a loss of high molecular weight VWF multimers. Recombinant VWF having the R273W mutation (rVWFR273W) expressed in COS-7 cells demonstrated severely impaired secretion and degradation in an intracellular location [Allen, S., et al. (2000) Blood 96, 560-568]. In this report we used pulse-chase analysis and endoglycosidase H digestion of wild-type rVWF and rVWFR273W immunoprecipitated from COS-7 cells to show that rVWFR273W was retained in the endoplasmic reticulum (ER). We demonstrate for the first time that wild-type rVWF and rVWFR273W interacted with the thiol-dependent oxidoreductase ERp57 during biosynthesis in the ER Pulse chase analysis demonstrated that the interactions of rVWFR273W with ERp57 and calnexin were prolonged compared to wild-type rVWF. In contrast there was no apparent difference between rVWFR273W and wild-type rVWF in their time-courses of interaction with calreticulin. (C) 2001 Academic Press.