Open AccessDesigner Palmitoylation Motif-Based Self-Localizing Ligand for Sustained Control of Protein Localization in Living Cells and Caenorhabditis elegans
Author(s) -
Akinobu Nakamura,
Choji Oki,
Shunsuke Sawada,
Tatsuyuki Yoshii,
Keiko Kuwata,
Andrew K. Rudd,
Neal K. Devaraj,
Kentaro Noma,
Shinya Tsukiji
Publication year - 2020
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.0c00014
Subject(s) - proteases , palmitoylation , caenorhabditis elegans , microbiology and biotechnology , biology , fusion protein , chromosomal translocation , cytoplasm , proteolysis , dihydrofolate reductase , mutant , biochemistry , enzyme , cysteine , gene , recombinant dna
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.