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Chemogenetic Control of Protein Anchoring to Endomembranes in Living Cells with Lipid-Tethered Small Molecules

preprint
submitted on 03.09.2019 and posted on 06.09.2019 by Akinobu Nakamura, Rika Katahira, Shunsuke Sawada, Eri Shinoda, Keiko Kuwata, Tatsuyuki Yoshii, Shinya Tsukiji
The Self-localizing Ligand-Induced Protein Translocation(SLIPT) system is an emerging platform that controls protein localization in living cells using synthetic self-localizing ligands (SLs). Here, we report a chemogenetic SLIPT system for inducing protein translocation from the cytoplasm to the surface of the endoplasmic reticulum (ER) and Golgi membranes, referred to as endomembranes.By screening a series of lipid-trimethoprim (TMP) conjugates, we found oleic acid-tethered TMP (oleTMP) to be the optimal SL that efficiently relocated and anchored Escherichiacoli dihydrofolate reductase (eDHFR)-fusion proteins toendomembranes. We showed that oleTMP mediated protein anchoring to endomembranes within minutes and could be reversed by the addition of free TMP. We also applied the endomembrane SLIPT system to artificially activate endomembrane Ras and inhibit the active nuclear transport of extracellular signal-regulated kinase (ERK), demonstrating its applicability for manipulating biological processes in living cells. We envision that the present oleTMP-based SLIPT system, which affords rapid and reversible control of protein anchoring to endomembranes, will offer a new unique tool for the study and control of spatiotemporally regulated cell signaling processes.

History

Email Address of Submitting Author

stsukiji@nitech.ac.jp

Institution

Nagoya Institute of Technology

Country

Japan

ORCID For Submitting Author

0000-0002-1402-5773

Declaration of Conflict of Interest

The authors declare no conflict of interests.

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