@article{06b90f3cc78e47eaae12864390802145,
title = "Multiplexed Delivery of Synthetic (Un)Conjugatable Ubiquitin and SUMO2 Enables Simultaneous Monitoring of Their Localization and Function in Live Cells",
abstract = "Ubiquitin (Ub) and its related small Ub like modifier (SUMO) are among the most influential protein post-translational modifications in eukaryotes. Unfortunately, visualizing these modifications in live cells is a challenging task. Chemical protein synthesis offers great opportunities in studying and further understanding Ub and SUMO biology. Nevertheless, the low cell permeability of proteins limits these studies mainly for in vitro applications. Here, we introduce a multiplexed protein cell delivery approach, termed MBL (multiplexed bead loading), for simultaneous loading of up to four differentially labeled proteins with organic fluorophores. We applied MBL to visualize ubiquitination and SUMOylation events in live and untransfected cells without fluorescent protein tags or perturbation to their endogenous levels. Our study reveals unprecedented involvements of Ub and SUMO2 in lysosomes depending on conjugation states. We envision that this approach will improve our understanding of dynamic cellular processes such as formation and disassembly of membraneless organelles.",
keywords = "chemical protein synthesis, fluorescent probes, post-translational modifications, protein delivery, protein modifications",
author = "Guy Mann and Pradeep Sadhu and Ashraf Brik",
note = "Funding Information: A.B. holds The Jordan and Irene Tark Academic Chair. This project has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement no. [831783]). We are grateful for Dr. Michael Lazarou of the Monash Biomedicine Discovery Institute for pBMN Parkin plasmid and for Dr. Michael H. Glickman and Prasad Sulkshane for establishing a stable +Parkin cell line with this plasmid. We are also grateful to Dr. Nitsan Dahan for his assistance in SIM2 imaging. The table of contents and Figure 7 were created using BioRender.com. Funding Information: A.B. holds The Jordan and Irene Tark Academic Chair. This project has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement no. [831783]). We are grateful for Dr. Michael Lazarou of the Monash Biomedicine Discovery Institute for pBMN Parkin plasmid and for Dr. Michael H. Glickman and Prasad Sulkshane for establishing a stable +Parkin cell line with this plasmid. We are also grateful to Dr. Nitsan Dahan for his assistance in SIM imaging. The table of contents and Figure 7 were created using BioRender.com. 2 Publisher Copyright: {\textcopyright} 2022 The Authors. ChemBioChem published by Wiley-VCH GmbH.",
year = "2022",
month = jun,
day = "3",
doi = "10.1002/cbic.202200122",
language = "English",
volume = "23",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley-VCH Verlag",
number = "11",
}