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) 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...
研究蛋白互作时，多种常规手段如酵母筛库、IP-MS、酵母双杂、Co-IP、BiFC、LCI等已经广为人知。今天，我将分享一种新近学到的技术：基于TurboID的邻近标记技术（Proximity Labeling, PL）。该技术在植物研究中尤为适用。PL技术的核心在于，通过融合具有连接酶活性的酶（如植物中的TurboID）和目标蛋白，催...
Proximity labeling techniques are emerging high-throughput methods for studying protein-protein, protein-RNA, and protein-DNA interactions with temporal and spatial precision. Proximity labeling methods take advantage of enzymes that can covalently label biomolecules with reactive substrates. These labeled ...
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
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...
191Citations 102Altmetric 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...
文献阅读-1:Exploiting effector-host interactions using TurboID-based proximity labeling 1只小脑斧ya 总要坚持些什么吧1 人赞同了该文章 科学问题:Ustilago maydis效应子UmSee1是玉米感染期间叶片中肿瘤形成所必需的。UmSee1与玉米SGT1相互作用并阻断其在体内的磷酸化。然而,目前尚不清楚哪些宿主过程被UmSee1和Um...
In the last two decades, different methods involving proximity labeling have been developed to study protein-protein interactions. In this review, we highlight the breadth and depth of these methods for the study of known protein-protein interactions as well as for the discovery of new protein in...