Direct Conjugation of NEDD8 to the N-Terminus of a Model Protein Can Induce Degradation.
Direct Conjugation of NEDD8 to the N-Terminus of a Model Protein Can Induce Degradation.
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NEDD8与模型蛋白的N末端的直接结合可以诱导降解。
DOI：
10.3390/cells10040854
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发表时间：
2021-04-09
期刊：
影响因子：
6
通讯作者：
Dolan BP
中科院分区：
文献类型：
--
作者：
Vijayasimha K;Tran MV;Leestemaker-Palmer AL;Dolan BP
虽然泛素在蛋白质降解中的作用已经很好地建立，但其他泛素样蛋白（UBL）在蛋白质降解中的作用尚不清楚。神经前体细胞表达的发育下调蛋白8（NEDD 8）是与泛素相比时鉴定的具有最高水平的氨基酸的UBL。在这里，我们测试了NEDD 8的N-末端添加到感兴趣的蛋白质是否会导致降解。正常NEDD 8加工所需的关键甘氨酸残基的突变导致不可切割的融合蛋白，其在细胞内被蛋白酶体和自噬快速降解。两种降解途径均依赖于功能性泛素结合系统，因为MLN 7243处理增加了不可裂解的NEDD 8-GFP水平。不可切割的N-末端NEDD 8-GFP的降解不是由于GFP折叠失败，因为具有不同折叠和荧光能力的不同NEDD 8-GFP构建体类似地降解。尽管NEDD 8与蛋白质的融合导致降解，但用MLN 4924（NEDD 8的E1激活酶抑制剂）处理细胞未能阻止其他不稳定底物的降解。总之，这些数据表明，在某些条件下，如本文所述的模型系统，NEDD 8与蛋白质底物的共价连接可能导致靶蛋白降解。
While the role of ubiquitin in protein degradation is well established, the role of other ubiquitin-like proteins (UBLs) in protein degradation is less clear. Neural precursor cell expressed developmentally down-regulated protein 8 (NEDD8) is the UBL with the highest level of amino acids identified when compared to ubiquitin. Here we tested if the N-terminal addition of NEDD8 to a protein of interest could lead to degradation. Mutation of critical glycine residues required for normal NEDD8 processing resulted in a non-cleavable fusion protein that was rapidly degraded within the cells by both the proteasome and autophagy. Both degradation pathways were dependent on a functional ubiquitin-conjugation system as treatment with MLN7243 increased levels of non-cleavable NEDD8-GFP. The degradation of non-cleavable, N-terminal NEDD8-GFP was not due to a failure of GFP folding as different NEDD8-GFP constructs with differing abilities to fold and fluoresce were similarly degraded. Though the fusion of NEDD8 to a protein resulted in degradation, treatment of cells with MLN4924, an inhibitor of the E1 activating enzyme for NEDD8, failed to prevent degradation of other destabilized substrates. Taken together these data suggest that under certain conditions, such as the model system described here, the covalent linkage of NEDD8 to a protein substrate may result in the target proteins degradation.
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发表时间：
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影响因子：
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发表时间：
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影响因子：
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发表时间：
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影响因子：
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