Designer Palmitoylation Motif-Based Self-Localizing Ligand for Sustained Control of Protein Localization in Living Cells and Caenorhabditis elegans

被引：20

作者：

Nakamura, Akinobu ^{[3
]}

Oki, Choji ^{[4
]}

Sawada, Shunsuke ^{[4
]}

Yoshii, Tatsuyuki ^{[3
,5
]}

Kuwata, Keiko ^{[6
]}

Rudd, Andrew K. ^{[7
]}

Devaraj, Neal K. ^{[7
]}

Noma, Kentaro ^{[5
,8
]}

Tsukiji, Shinya ^{[1
,2
]}

机构：

[1] Nagoya Inst Technol, Dept Life Sci & Appl Chem, Dept Nanopharmaceut Sci, Nagoya, Aichi 4668555, Japan

[2] Nagoya Inst Technol, FRIMS, Nagoya, Aichi 4668555, Japan

[3] Nagoya Inst Technol, Dept Life Sci & Appl Chem, Nagoya, Aichi 4668555, Japan

[4] Nagoya Inst Technol, Dept Nanopharmaceut Sci, Nagoya, Aichi 4668555, Japan

[5] Japan Sci & Technol Agcy JST, PRESTO, Saitama 3320012, Japan

[6] Nagoya Univ, Inst Transformat Biomol ITbM, Nagoya, Aichi 4648602, Japan

[7] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92093 USA

[8] Nagoya Univ, Neurosci Inst, Grad Sch Sci, Grp Nutr Neurosci, Nagoya, Aichi 4648602, Japan

来源：

ACS CHEMICAL BIOLOGY | 2020年 / 15卷 / 04期

基金：

日本科学技术振兴机构;

关键词：

D-AMINO-ACID; MEMBRANE; ACTIVATION; ERK; SUBSTITUTION; STABILITY; RAF;

D O I：

10.1021/acschembio.0c00014

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Inducing protein translocation to the plasma membrane (PM) is an important approach for manipulating diverse signaling molecules/pathways in living cells. We previously devised a new chemogenetic system, in which a protein fused to Escherichia coli dihydrofolate reductase (eDHFR) can be rapidly translocated from the cytoplasm to the PM using a trimethoprim (TMP)-based self-localizing ligand (SL), mgcTMP. However, mgcTMP-induced protein translocation turned out to be transient and spontaneously reversed within 1 h, limiting its application. Here, we first demonstrated that the spontaneous reverse translocation was caused by cellular degradation of mgcTMP, presumably by proteases. To address this problem, we newly developed a proteolysis-resistant SL, m(D) cTMP. This m(D) cTMP now allows sustained PM localization of eDHFR -fusion proteins (over several hours to a day), and it was applicable to inducing prolonged signal activation and cell differentiation . m(D) cTMP also worked in live nematodes, making it an attractive new tool for probing and controlling living systems.

引用

页码：837 / 843

页数：7