邻近标记(proximity labeling, PL)技术的基本原理是将一个具有连接酶活性的酶(目前植物中广泛应用的为TurboID)与目的蛋白融合, 在连接酶的催化作用下将小分子底物(如生物素(biotin))连接到与目的蛋白邻近的内源蛋白或RNA, 而后可通过富集被标记的蛋白或RNA分别进行质谱分析或RNA-Seq, 可鉴定目的蛋白邻近的蛋白或RNA...
Proximity Labeling(PL),邻近标记 原理 将一个具有邻近标记功能的酶(PL ligase)与目标蛋白(诱饵蛋白,Bait protein)融合,通过酶催化的共价修饰将邻近的蛋白标记上生物素(biotin),最后通过亲和素磁珠富集生物素标记蛋白进行质谱鉴定,分析目的蛋白的互作或邻近的蛋白信息。 Mathew B, et al. Mol Cell Proteomics. (202...
Proximity labeling (PL) 临近标记检测临近标记法互作蛋白检测(Proximity labeling,PL) 临近标记法互作蛋白检测(Proximity labeling,PL),是一种通过融合外源的酶基因到诱饵蛋白基因,然后催化生成可扩散的小分子,来标记细胞内源性的临...查看详细>>关于怡科
临近标记法互作蛋白检测(Proximity labeling,PL),是一种通过融合外源的酶基因到诱饵蛋白基因,然后催化生成可扩散的小分子,来标记细胞内源性的临近蛋白质,然后通过富集、高分辨率质谱或核酸测序来检测鉴定与靶蛋白互作的蛋白质与核酸。相比传统免疫共沉淀(co-IP)与pull down法,临近标记法检测互作蛋白的最大优势就是能...
Using a proximity-dependent labeling system, we identified distinct classes of proteins that regulate R-loops in vivo through different mechanisms. We show that ATRX suppresses R-loops by interacting with RNAs and preventing R-loop formation. Our proteomics screen also discovered an unexpected ...
184Citations 107Altmetric Metrics Abstract This protocol describes the use of TurboID and split-TurboID in proximity labeling applications for mapping protein–protein interactions and subcellular proteomes in live mammalian cells. TurboID is an engineered biotin ligase that uses ATP to convert biotin int...
This protocol describes the use of TurboID and split-TurboID in proximity labeling applications for mapping protein–protein interactions and subcellular proteomes in live mammalian cells. TurboID is an engineered biotin ligase that uses ATP to convert biotin into biotin–AMP, a reactive intermediate th...
Proximity LabelingMurat Sunbul Andres Jäschke EditorsMethods and ProtocolsMethods in Molecular Biology 2008 阅读了该文档的用户还阅读了这些文档 108 p. ISO 25539-3-2024 18 p. ISO 16810-2024 38 p. ISO 14356-2024 10 p. ISO 15708-1-2024 72 p. BS EN 1982-2024 50 p. PD ISO-...
TurboID邻近标记(Proximity labeling, PL)的原理是将TurboID生物素连接酶与诱饵蛋白融合,当诱饵蛋白与靶标蛋白发生互作时,TurboID生物素连接酶通过催化的共价修饰将邻近的蛋白标记上生物素,最后通过亲和素磁珠富集生物素标记的蛋白进行质谱鉴定,从而获得靶标蛋白信息。
Proximity-labeling-based proteomic approaches have been recently applied to plant research and have been shown to be invaluable in profiling functional components of various protein complexes10,11,12,13,14. By combining the power of BioID2-based proximity labeling (PL)15, label-free quantitative ma...