Designer Palmitoylation Motif-Based Self-Localizing Ligand for Sustained Control of Protein Localization in Living Cells and Caenorhabditis elegans
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 tran...
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| Veröffentlicht in: | ACS chemical biology 2020-04, Vol.15 (4), p.837-843 |
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| creator | Nakamura, Akinobu Oki, Choji Sawada, Shunsuke Yoshii, Tatsuyuki Kuwata, Keiko Rudd, Andrew K Devaraj, Neal K Noma, Kentaro Tsukiji, Shinya |
| description | 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, mDcTMP. This mDcTMP 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. mDcTMP also worked in live nematodes, making it an attractive new tool for probing and controlling living systems. |
| doi_str_mv | 10.1021/acschembio.0c00014 |
| format | Article |
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Biol</addtitle><description>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, mDcTMP. 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Oki, Choji ; Sawada, Shunsuke ; Yoshii, Tatsuyuki ; Kuwata, Keiko ; Rudd, Andrew K ; Devaraj, Neal K ; Noma, Kentaro ; Tsukiji, Shinya</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a441t-8adfa88777db45996f24684ddabaea2b550ecc509922e6f6f96c3543a980e20c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Caenorhabditis elegans</topic><topic>Caenorhabditis elegans Proteins - metabolism</topic><topic>Cell Line, Tumor</topic><topic>Cell Membrane - metabolism</topic><topic>Cysteine - analogs & derivatives</topic><topic>Cysteine - metabolism</topic><topic>Cysteine - pharmacology</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli Proteins - metabolism</topic><topic>Golgi Apparatus - metabolism</topic><topic>Humans</topic><topic>Ligands</topic><topic>Lipoylation</topic><topic>Protein Transport - drug effects</topic><topic>Proto-Oncogene Proteins c-raf - metabolism</topic><topic>Rats</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Signal Transduction - physiology</topic><topic>Stereoisomerism</topic><topic>Tetrahydrofolate Dehydrogenase - metabolism</topic><topic>Trimethoprim - analogs & derivatives</topic><topic>Trimethoprim - metabolism</topic><topic>Trimethoprim - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nakamura, Akinobu</creatorcontrib><creatorcontrib>Oki, Choji</creatorcontrib><creatorcontrib>Sawada, Shunsuke</creatorcontrib><creatorcontrib>Yoshii, Tatsuyuki</creatorcontrib><creatorcontrib>Kuwata, Keiko</creatorcontrib><creatorcontrib>Rudd, Andrew K</creatorcontrib><creatorcontrib>Devaraj, Neal K</creatorcontrib><creatorcontrib>Noma, Kentaro</creatorcontrib><creatorcontrib>Tsukiji, Shinya</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>ACS chemical biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nakamura, Akinobu</au><au>Oki, Choji</au><au>Sawada, Shunsuke</au><au>Yoshii, Tatsuyuki</au><au>Kuwata, Keiko</au><au>Rudd, Andrew K</au><au>Devaraj, Neal K</au><au>Noma, Kentaro</au><au>Tsukiji, Shinya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Designer Palmitoylation Motif-Based Self-Localizing Ligand for Sustained Control of Protein Localization in Living Cells and Caenorhabditis elegans</atitle><jtitle>ACS chemical biology</jtitle><addtitle>ACS Chem. 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| ispartof | ACS chemical biology, 2020-04, Vol.15 (4), p.837-843 |
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| source | ACS Publications; MEDLINE |
| subjects | Animals Caenorhabditis elegans Caenorhabditis elegans Proteins - metabolism Cell Line, Tumor Cell Membrane - metabolism Cysteine - analogs & derivatives Cysteine - metabolism Cysteine - pharmacology Escherichia coli - enzymology Escherichia coli Proteins - metabolism Golgi Apparatus - metabolism Humans Ligands Lipoylation Protein Transport - drug effects Proto-Oncogene Proteins c-raf - metabolism Rats Recombinant Fusion Proteins - metabolism Signal Transduction - physiology Stereoisomerism Tetrahydrofolate Dehydrogenase - metabolism Trimethoprim - analogs & derivatives Trimethoprim - metabolism Trimethoprim - pharmacology |
| title | Designer Palmitoylation Motif-Based Self-Localizing Ligand for Sustained Control of Protein Localization in Living Cells and Caenorhabditis elegans |
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