A proximity labeling protocol to probe proximity interactions in C. elegans.
A proximity labeling protocol to probe proximity interactions in C. elegans.
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DOI：
10.1016/j.xpro.2021.100986
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发表时间：
2021-12-17
期刊：
影响因子：
--
通讯作者：
Feldman JL
中科院分区：
文献类型：
--
作者：
Sanchez AD;Feldman JL
酶催化邻近标记（PL）已成为识别细胞中蛋白质-蛋白质邻近相互作用的关键方法;然而，由于技术限制，PL技术在活的多细胞生物体中历史上并不实用。在这里，我们提出了一个协议应用PL活线虫使用生物素连接酶突变酶TurboID。我们通过关注肠细胞的非中心体MTOC（ncMTOC），以组织特异性和区域特异性的方式证明了PL。该方案可用于靶向体内蛋白质网络分析。有关本方案使用和执行的完整详细信息，请参阅。基于TurboID的秀丽隐杆线虫生物素邻近标记实验设计指南从转基因设计到蛋白质鉴定的逐步TurboID实验酶催化邻近标记（PL）已成为鉴定细胞中蛋白质-蛋白质邻近相互作用的关键方法;然而，由于技术限制，PL技术在活的多细胞生物体中历史上并不实用。在这里，我们提出了一个协议应用PL活线虫使用生物素连接酶突变酶TurboID。我们通过关注肠细胞的非中心体MTOC（ncMTOC），以组织特异性和区域特异性的方式证明了PL。该方案可用于靶向体内蛋白质网络分析。
Enzyme-catalyzed proximity labeling (PL) has emerged as a critical approach for identifying protein-protein proximity interactions in cells; however, PL techniques were not historically practical in living multicellular organisms due to technical limitations. Here, we present a protocol for applying PL to living C. elegans using the biotin ligase mutant enzyme TurboID. We demonstrated PL in a tissue-specific and region-specific manner by focusing on non-centrosomal MTOCs (ncMTOCs) of intestinal cells. This protocol is useful for targeted in vivo protein network profiling. For complete details on the use and execution of this protocol, please refer to. TurboID enabled biotin-based proximity labeling protocol for C. elegans Experimental design guidelines for proximity labeling in C. elegans A step-by-step TurboID protocol from transgene design to protein identification Enzyme-catalyzed proximity labeling (PL) has emerged as a critical approach for identifying protein-protein proximity interactions in cells; however, PL techniques were not historically practical in living multicellular organisms due to technical limitations. Here, we present a protocol for applying PL to living C. elegans using the biotin ligase mutant enzyme TurboID. We demonstrated PL in a tissue-specific and region-specific manner by focusing on non-centrosomal MTOCs (ncMTOCs) of intestinal cells. This protocol is useful for targeted in vivo protein network profiling.
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影响因子：
14.8
作者：
Hung V;Udeshi ND;Lam SS;Loh KH;Cox KJ;Pedram K;Carr SA;Ting AY
通讯作者：
Ting AY
DOI：
10.1016/j.cub.2021.06.021
发表时间：
2021-08-23
期刊：
Current biology : CB
影响因子：
--
作者：
Sanchez AD;Branon TC;Cote LE;Papagiannakis A;Liang X;Pickett MA;Shen K;Jacobs-Wagner C;Ting AY;Feldman JL
通讯作者：
Feldman JL
影响因子：
7.7
作者：
Wang S;Wu D;Quintin S;Green RA;Cheerambathur DK;Ochoa SD;Desai A;Oegema K
通讯作者：
Oegema K
使用 TurboID 在活细胞和生物体中进行高效邻近标记。
影响因子：
46.9
作者：
Branon TC;Bosch JA;Sanchez AD;Udeshi ND;Svinkina T;Carr SA;Feldman JL;Perrimon N;Ting AY
通讯作者：
Ting AY
影响因子：
7.7
作者：
Richardson CE;Spilker KA;Cueva JG;Perrino J;Goodman MB;Shen K
通讯作者：
Shen K