| RRC ID |
73107
|
| 著者 |
Suzuki S, Nakamura A, Hatano Y, Yoshikawa M, Yoshii T, Sawada S, Atsuta-Tsunoda K, Aoki K, Tsukiji S.
|
| タイトル |
A chemogenetic platform for controlling plasma membrane signaling and synthetic signal oscillation.
|
| ジャーナル |
Cell Chem Biol
|
| Abstract |
Chemogenetic methods enabling the rapid translocation of specific proteins to the plasma membrane (PM) in a single protein-single ligand manner are useful tools in cell biology. We recently developed a technique, in which proteins fused to an Escherichia coli dihydrofolate reductase (eDHFR) variant carrying N-terminal hexalysine residues are recruited from the cytoplasm to the PM using the synthetic myristoyl-d-Cys-tethered trimethoprim (mDcTMP) ligand. However, this system achieved PM-specific translocation only when the eDHFR tag was fused to the N terminus of proteins, thereby limiting its application. In this report, we engineered a universal PM-targeting tag for mDcTMP-induced protein translocation by grafting the hexalysine motif into an intra-loop region of eDHFR. We demonstrate the broad applicability of the new loop-engineered eDHFR tag and mDcTMP pair for conditional PM recruitment and activation of various tag-fused signaling proteins with different fusion configurations and for reversibly and repeatedly controlling protein localization to generate synthetic signal oscillations.
|
| 巻・号 |
29(9)
|
| ページ |
1446-1464.e10
|
| 公開日 |
2022-9-15
|
| DOI |
10.1016/j.chembiol.2022.06.005
|
| PII |
S2451-9456(22)00237-9
|
| PMID |
35835118
|
| MeSH |
Cell Membrane / metabolism
Escherichia coli / metabolism
Ligands
Proteins
Signal Transduction
Tetrahydrofolate Dehydrogenase* / metabolism
Trimethoprim* / pharmacology
|
| IF |
6.762
|
| リソース情報 |
| 遺伝子材料 |
pCMV-VSV-G-RSV-Re (RDB04393) |
