A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
Bioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an...
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| Format: | Article |
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Frontiers Media S.A.
2021-11-01
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| Series: | Frontiers in Chemistry |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fchem.2021.768535/full |
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| author | Birthe Meineke Birthe Meineke Johannes Heimgärtner Johannes Heimgärtner Alexander J. Craig Michael Landreh Lindon W. K. Moodie Lindon W. K. Moodie Simon J. Elsässer Simon J. Elsässer |
| author_facet | Birthe Meineke Birthe Meineke Johannes Heimgärtner Johannes Heimgärtner Alexander J. Craig Michael Landreh Lindon W. K. Moodie Lindon W. K. Moodie Simon J. Elsässer Simon J. Elsässer |
| author_sort | Birthe Meineke |
| collection | DOAJ |
| description | Bioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an efficient method for incorporating such chemical handles into proteins on the ribosome, in which noncanonical amino acids (ncAAs) are site specifically introduced into the polypeptide in response to an amber (UAG) stop codon. A variety of ncAA structures containing azides or alkynes have been proven useful for performing CuAAC chemistry on proteins. To improve CuAAC efficiency, biologically incorporated alkyne groups can be reacted with azide substrates that contain copper-chelating groups. However, the direct incorporation of copper-chelating azides into proteins has not been explored. To remedy this, we prepared the ncAA paz-lysine (PazK), which contains a picolyl azide motif. We show that PazK is efficiently incorporated into proteins by amber suppression in mammalian cells. Furthermore, PazK-labeled proteins show improved reactivity with alkyne reagents in CuAAC. |
| first_indexed | 2024-12-19T00:18:16Z |
| format | Article |
| id | doaj.art-80dfbc6fdcfb4fc3aa01db18bc5f6838 |
| institution | Directory Open Access Journal |
| issn | 2296-2646 |
| language | English |
| last_indexed | 2024-12-19T00:18:16Z |
| publishDate | 2021-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Chemistry |
| spelling | doaj.art-80dfbc6fdcfb4fc3aa01db18bc5f68382022-12-21T20:45:41ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462021-11-01910.3389/fchem.2021.768535768535A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click LabelingBirthe Meineke0Birthe Meineke1Johannes Heimgärtner2Johannes Heimgärtner3Alexander J. Craig4Michael Landreh5Lindon W. K. Moodie6Lindon W. K. Moodie7Simon J. Elsässer8Simon J. Elsässer9Science for Life Laboratory, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Stockholm, SwedenMing Wai Lau Centre for Reparative Medicine, Stockholm Node, Karolinska Institutet, Stockholm, SwedenScience for Life Laboratory, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Stockholm, SwedenMing Wai Lau Centre for Reparative Medicine, Stockholm Node, Karolinska Institutet, Stockholm, SwedenDrug Design and Discovery, Department of Medicinal Chemistry, Biomedical Centre, Uppsala University, Uppsala, SwedenDepartment of Microbiology, Tumor and Cell Biology, Science for Life Laboratory, Karolinska Institutet, Stockholm, SwedenDrug Design and Discovery, Department of Medicinal Chemistry, Biomedical Centre, Uppsala University, Uppsala, SwedenUppsala Antibiotic Centre, Uppsala University, Uppsala, SwedenScience for Life Laboratory, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Stockholm, SwedenMing Wai Lau Centre for Reparative Medicine, Stockholm Node, Karolinska Institutet, Stockholm, SwedenBioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an efficient method for incorporating such chemical handles into proteins on the ribosome, in which noncanonical amino acids (ncAAs) are site specifically introduced into the polypeptide in response to an amber (UAG) stop codon. A variety of ncAA structures containing azides or alkynes have been proven useful for performing CuAAC chemistry on proteins. To improve CuAAC efficiency, biologically incorporated alkyne groups can be reacted with azide substrates that contain copper-chelating groups. However, the direct incorporation of copper-chelating azides into proteins has not been explored. To remedy this, we prepared the ncAA paz-lysine (PazK), which contains a picolyl azide motif. We show that PazK is efficiently incorporated into proteins by amber suppression in mammalian cells. Furthermore, PazK-labeled proteins show improved reactivity with alkyne reagents in CuAAC.https://www.frontiersin.org/articles/10.3389/fchem.2021.768535/fullgenetic code expansionamber suppressionnoncanonical amino acidbioorthogonal chemistryclick chemistrycopper catalyzed azide–alkyne cycloaddition (CuAAC) |
| spellingShingle | Birthe Meineke Birthe Meineke Johannes Heimgärtner Johannes Heimgärtner Alexander J. Craig Michael Landreh Lindon W. K. Moodie Lindon W. K. Moodie Simon J. Elsässer Simon J. Elsässer A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling Frontiers in Chemistry genetic code expansion amber suppression noncanonical amino acid bioorthogonal chemistry click chemistry copper catalyzed azide–alkyne cycloaddition (CuAAC) |
| title | A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling |
| title_full | A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling |
| title_fullStr | A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling |
| title_full_unstemmed | A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling |
| title_short | A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling |
| title_sort | genetically encoded picolyl azide for improved live cell copper click labeling |
| topic | genetic code expansion amber suppression noncanonical amino acid bioorthogonal chemistry click chemistry copper catalyzed azide–alkyne cycloaddition (CuAAC) |
| url | https://www.frontiersin.org/articles/10.3389/fchem.2021.768535/full |
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