bi9b00807_si_003.mov (38.25 MB)
Chemogenetic Control of Protein Anchoring to Endomembranes in Living Cells with Lipid-Tethered Small Molecules
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posted on 2019-10-15, 14:46 authored by Akinobu Nakamura, Rika Katahira, Shunsuke Sawada, Eri Shinoda, Keiko Kuwata, Tatsuyuki Yoshii, Shinya TsukijiThe
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 Escherichia coli dihydrofolate reductase (eDHFR)-fusion
proteins to endomembranes. 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.
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Chemogenetic Controlendomembrane Rascontrols protein localizationendomembrane SLIPT systemoleic acid-tethered TMPGolgi membranesendoplasmic reticulumERKextracellular signal-regulated kinaseLiving CellsoleTMP-based SLIPT systemself-localizing ligandsprotein translocationProtein AnchoringERself-localizing ligand-induced protein translocationchemogenetic SLIPT systemEscherichia coli dihydrofolate reductase
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