修剪
泛素
细胞生物学
化学
HEK 293细胞
泛素蛋白连接酶类
泛素连接酶
生物
计算机科学
生物化学
基因
操作系统
作者
Tycho E.T. Mevissen,Anisa V. Prasad,Johannes C. Walter
出处
期刊:Cell Reports
[Elsevier]
日期:2023-02-01
卷期号:42 (2): 112125-112125
被引量:6
标识
DOI:10.1016/j.celrep.2023.112125
摘要
Tripartite motif-containing protein 21 (TRIM21) is a cytosolic antibody receptor and E3 ubiquitin ligase that promotes destruction of a broad range of pathogens. TRIM21 also underlies the antibody-dependent protein targeting method Trim-Away. Current evidence suggests that TRIM21 binding to antibodies leads to formation of a self-anchored K63 ubiquitin chain on the N terminus of TRIM21 that triggers the destruction of TRIM21, antibody, and target protein. Here, we report that addition of antibody and TRIM21 to Xenopus egg extracts promotes efficient degradation of endogenous target proteins, establishing cell-free Trim-Away as a powerful tool to interrogate protein function. Chemical methylation of TRIM21 had no effect on target proteolysis, whereas deletion of all lysine residues in targets abolished their ubiquitination and proteasomal degradation. These results demonstrate that target protein, but not TRIM21, polyubiquitination is required for Trim-Away, and they suggest that current models of TRIM21 function should be fundamentally revised.
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