However, current PL methods require over 18 h of labeling time or utilize chemicals with limited cell permeability or high toxicity. 2012;196:801–810. -, Choi-Rhee E, Schulman H, Cronan JE. Meyer M, Ryck J, Goormachtig S, Van Damme P. Int J Mol Sci. Proximity labeling catalyzed by promiscuous enzymes, such as TurboID, have enabled the proteomic analysis of subcellular regions difficult or impossible to access by conventional fractionation-based approaches. | and A.Y.T. Author Correction: Efficient proximity labeling in living cells and organisms with TurboID. HHS Science. We need a deliberative process that’s transparent, politically neutral, scientifically balanced, and representative of all relevant disciplines and expertise. doi: 10.7554/eLife.47864. NOTE: Your email address is requested solely to identify you as the sender of this article. Get the latest public health information from CDC: https://www.coronavirus.gov. We used yeast display-based directed evolution to engineer two promiscuous mutants of biotin ligase, TurboID and miniTurbo, which catalyze PL with much greater efficiency than BioID or BioID2, and enable 10-min PL in cells with non-toxic and easily deliverable biotin. | Impact Statement TurboID-mediated proximity labeling is a powerful tool for protein interactomics in plants. -, Rhee HW, et al. Yet some cellular regions, such as organelle contact sites, remain out of reach for current PL methods. Here we demonstrate the application of enzyme-catalyzed proximity labeling (PL), using the engineered promiscuous biotin ligase TurboID, as a sensitive method for characterizing PPIs in signaling networks. Zhang Y, Li Y, Yang X, Wen Z, Nagalakshmi U, Dinesh-Kumar SP. Trends in Cell Biology. However, low kinetics and the requirement of toxic chemicals limit the broad utilisation of current proximity labelling methods in living organisms. DP2 GM119136/GM/NIGMS NIH HHS/United States, R01 CA186568/CA/NCI NIH HHS/United States, T32 GM007276/GM/NIGMS NIH HHS/United States, U24 CA210986/CA/NCI NIH HHS/United States, Kim DI, Roux KJ. A.Y.T. wrote the paper. designed research; K.F.C., T.C.B., S.R., T.S., N.D.U., T.T., C.K., H.-W.R., and I.-K.L. Protein Sci. 2013;339:1328–1331. Copyright © 2020 National Academy of Sciences. analyzed data; and K.F.C. Find NCBI SARS-CoV-2 literature, sequence, and clinical content: https://www.ncbi.nlm.nih.gov/sars-cov-2/. You may purchase access to this article. Epub 2016 Jul 27. Characterization of TurboID and miniTurbo…, Figure 2. Yeow ZY, Lambrus BG, Marlow R, Zhan KH, Durin MA, Evans LT, Scott PM, Phan T, Park E, Ruiz LA, Moralli D, Knight EG, Badder LM, Novo D, Haider S, Green CM, Tutt ANJ, Lord CJ, Chapman JR, Holland AJ. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We optimized the protocol of biotin labeling and affinity purification in transgenic Arabidopsis expressing a BIN2-TurboID fusion protein. The authors declare no competing interest. Enter multiple addresses on separate lines or separate them with commas. Subsequent quantitative mass spectrometry (MS) analysis identified about three hundred proteins biotinylated by BIN2-TurboID more efficiently than the YFP-TurboID control.
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