Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells
To enable minimally invasive studies of proteins in their native context, it is desirable to tag proteins with small, bright reporter groups. Recently, our lab described PRIME technology (for PRobe Incorporation Mediated by Enzymes) for such tagging[1-3]. An engineered variant of Escherichia coli li...
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| Format: | Article |
| Language: | en_US |
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Wiley-VCH Verlag
2012
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| Online Access: | http://hdl.handle.net/1721.1/69559 https://orcid.org/0000-0002-8277-5226 |
| _version_ | 1826204370242371584 |
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| author | Jin, Xin Uttamapinant, Chayasith Ting, Alice Y. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Jin, Xin Uttamapinant, Chayasith Ting, Alice Y. |
| author_sort | Jin, Xin |
| collection | MIT |
| description | To enable minimally invasive studies of proteins in their native context, it is desirable to tag proteins with small, bright reporter groups. Recently, our lab described PRIME technology (for PRobe Incorporation Mediated by Enzymes) for such tagging[1-3]. An engineered variant of Escherichia coli lipoic acid ligase (LplA) is used to covalently attach a fluorescent substrate, such as 7-hydroxycoumarin, onto a 13-amino acid peptide recognition sequence (called LAP, for Ligase Acceptor Peptide) that is genetically fused to a protein of interest (POI) (Figure 1A). The targeting specificity is derived from the extremely high natural sequence specificity of LplA[4]. PRIME was used to label and visualize various LAP-tagged cytoskeletal and adhesion proteins in living mammalian cells. PRIME time: We report the synthesis of a pH-insensitive blue fluorophore, 7-aminocoumarin, by using palladium- catalyzed Buchwald–Hartwig cross coupling. 7-Aminocoumarin can be used to tag recombinant proteins on the cell surface and inside living cells through PRIME (probe incorporation mediated by enzymes), and unlike 7-hydroxycoumarin, can be visualized in acidic organelles such as endosomes. |
| first_indexed | 2024-09-23T12:53:34Z |
| format | Article |
| id | mit-1721.1/69559 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:53:34Z |
| publishDate | 2012 |
| publisher | Wiley-VCH Verlag |
| record_format | dspace |
| spelling | mit-1721.1/695592022-09-28T10:42:16Z Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells Jin, Xin Uttamapinant, Chayasith Ting, Alice Y. Massachusetts Institute of Technology. Department of Chemistry Ting, Alice Y. Ting, Alice Y. Jin, Xin Uttamapinant, Chayasith To enable minimally invasive studies of proteins in their native context, it is desirable to tag proteins with small, bright reporter groups. Recently, our lab described PRIME technology (for PRobe Incorporation Mediated by Enzymes) for such tagging[1-3]. An engineered variant of Escherichia coli lipoic acid ligase (LplA) is used to covalently attach a fluorescent substrate, such as 7-hydroxycoumarin, onto a 13-amino acid peptide recognition sequence (called LAP, for Ligase Acceptor Peptide) that is genetically fused to a protein of interest (POI) (Figure 1A). The targeting specificity is derived from the extremely high natural sequence specificity of LplA[4]. PRIME was used to label and visualize various LAP-tagged cytoskeletal and adhesion proteins in living mammalian cells. PRIME time: We report the synthesis of a pH-insensitive blue fluorophore, 7-aminocoumarin, by using palladium- catalyzed Buchwald–Hartwig cross coupling. 7-Aminocoumarin can be used to tag recombinant proteins on the cell surface and inside living cells through PRIME (probe incorporation mediated by enzymes), and unlike 7-hydroxycoumarin, can be visualized in acidic organelles such as endosomes. National Institutes of Health (U.S.) (NIH (R01 GM072670)) Massachusetts Institute of Technology (John Reed (MIT Class of 1961)) Massachusetts Institute of Technology (Paul E. Gray (MIT Class of 1954)) 2012-03-02T15:55:50Z 2012-03-02T15:55:50Z 2011-01 2010-07 Article http://purl.org/eprint/type/JournalArticle 1439-4227 1439-7633 http://hdl.handle.net/1721.1/69559 Jin, Xin, Chayasith Uttamapinant, and Alice Y. Ting. “Synthesis of 7-Aminocoumarin by Buchwald-Hartwig Cross Coupling for Specific Protein Labeling in Living Cells.” ChemBioChem 12.1 (2011): 65–70. https://orcid.org/0000-0002-8277-5226 en_US http://dx.doi.org/10.1002/cbic.201000414 ChemBioChem Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf Wiley-VCH Verlag Prof. Ting via Erja Kajosalo |
| spellingShingle | Jin, Xin Uttamapinant, Chayasith Ting, Alice Y. Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells |
| title | Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells |
| title_full | Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells |
| title_fullStr | Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells |
| title_full_unstemmed | Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells |
| title_short | Synthesis of 7-aminocoumarin via Buchwald-Hartwig cross coupling for specific protein labeling in living cells |
| title_sort | synthesis of 7 aminocoumarin via buchwald hartwig cross coupling for specific protein labeling in living cells |
| url | http://hdl.handle.net/1721.1/69559 https://orcid.org/0000-0002-8277-5226 |
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