Identification of ERAD components essential for dislocation of the null Hong Kong variant of α-1-antitrypsin (NHK)
Identification of ERAD components essential for dislocation of the null Hong Kong variant of α-1-antitrypsin (NHK)
复制标题
鉴定对于 α-1-抗胰蛋白酶香港无效变体脱位至关重要的 ERAD 成分 (NHK)
DOI：
10.1016/j.bbrc.2015.01.133
复制
发表时间：
2015
影响因子：
3.1
通讯作者：
Fang, Shengyun
中科院分区：
文献类型：
--
作者：
Zhong, Yongwang;Shen, Hang;Wang, Ye;Yang, Yili;Yang, Peixin;Fang, Shengyun
错误折叠的蛋白质或蛋白质复合物的孤儿亚基通过 ER 相关降解 (ERAD) 从内质网 (ER) 中去除。 ERAD 需要将那些不需要的蛋白质从 ER 腔错位（也称为逆转位）到胞质溶胶，以便被蛋白酶体破坏。超过一百种 ERAD 成分蛋白已被鉴定，但它们在错位中的作用仍知之甚少。在这里，我们使用我们最近开发的位错诱导重组 GFP (drGFP) 测定法评估了活细胞中 NHK 错位的 ERAD 成分的需求。 RNAi显示，所检查的21个ERAD成分中有12个是NHK有效脱位所必需的，其中Hrd1、Sel1L、GRP94和p97/VCP是至关重要的。此外，敲除 21 个组件中的 7 个会增强 NHK 的错位。这项研究揭示了 NHK 错位所需的复杂蛋白质功能网络。
Misfolded proteins or orphan subunits of protein complexes are removed from the endoplasmic reticulum (ER) by ER-associated degradation (ERAD). ERAD requires dislocation, also known as retrotranslocation, of those unwanted proteins from the ER lumen to the cytosol for destruction by the proteasomes. Over one hundred ERAD component proteins have been identified but their role in dislocation remain poorly understood. Here we assessed the requirement of ERAD components for dislocation of NHK in live cells using our recently developed dislocation-induced reconstituted GFP (drGFP) assay. RNAi revealed that 12 out of 21 ERAD components examined are required for efficient dislocation of NHK among which Hrd1, Sel1L, GRP94 and p97/VCP are critically required. In addition, knockdown of 7 of the 21 components enhanced NHK dislocation. This study uncovers a complex functional network of proteins required for NHK dislocation.