Site-Selective Cysteine-Cyclooctyne Conjugation
We report a site-selective cysteine–cyclooctyne conjugation reaction between a seven-residue peptide tag (DBCO-tag, Leu-Cys-Tyr-Pro-Trp-Val-Tyr) at the N or C terminus of a peptide or protein and various aza-dibenzocyclooctyne (DBCO) reagents. Compared to a cysteine peptide control, the DBCO-tag inc...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2020
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| Online Access: | https://hdl.handle.net/1721.1/123516 |
| _version_ | 1826190902745366528 |
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| author | Zhang, Chi Dai, Peng Vinogradov, Alexander A. Gates, Zachary P Pentelute, Bradley L. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Zhang, Chi Dai, Peng Vinogradov, Alexander A. Gates, Zachary P Pentelute, Bradley L. |
| author_sort | Zhang, Chi |
| collection | MIT |
| description | We report a site-selective cysteine–cyclooctyne conjugation reaction between a seven-residue peptide tag (DBCO-tag, Leu-Cys-Tyr-Pro-Trp-Val-Tyr) at the N or C terminus of a peptide or protein and various aza-dibenzocyclooctyne (DBCO) reagents. Compared to a cysteine peptide control, the DBCO-tag increases the rate of the thiol–yne reaction 220-fold, thereby enabling selective conjugation of DBCO-tag to DBCO-linked fluorescent probes, affinity tags, and cytotoxic drug molecules. Fusion of DBCO-tag with the protein of interest enables regioselective cysteine modification on proteins that contain multiple endogenous cysteines; these examples include green fluorescent protein and the antibody trastuzumab. This study demonstrates that short peptide tags can aid in accelerating bond-forming reactions that are often slow to non-existent in water. Keywords: bioconjugation; bioorthogonal chemistry; cysteine–cyclooctyne reaction; dibenzocyclooctyne, protein modification |
| first_indexed | 2024-09-23T08:47:05Z |
| format | Article |
| id | mit-1721.1/123516 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T08:47:05Z |
| publishDate | 2020 |
| publisher | Wiley |
| record_format | dspace |
| spelling | mit-1721.1/1235162022-09-23T14:32:01Z Site-Selective Cysteine-Cyclooctyne Conjugation Zhang, Chi Dai, Peng Vinogradov, Alexander A. Gates, Zachary P Pentelute, Bradley L. Massachusetts Institute of Technology. Department of Chemistry We report a site-selective cysteine–cyclooctyne conjugation reaction between a seven-residue peptide tag (DBCO-tag, Leu-Cys-Tyr-Pro-Trp-Val-Tyr) at the N or C terminus of a peptide or protein and various aza-dibenzocyclooctyne (DBCO) reagents. Compared to a cysteine peptide control, the DBCO-tag increases the rate of the thiol–yne reaction 220-fold, thereby enabling selective conjugation of DBCO-tag to DBCO-linked fluorescent probes, affinity tags, and cytotoxic drug molecules. Fusion of DBCO-tag with the protein of interest enables regioselective cysteine modification on proteins that contain multiple endogenous cysteines; these examples include green fluorescent protein and the antibody trastuzumab. This study demonstrates that short peptide tags can aid in accelerating bond-forming reactions that are often slow to non-existent in water. Keywords: bioconjugation; bioorthogonal chemistry; cysteine–cyclooctyne reaction; dibenzocyclooctyne, protein modification National Institute of General Medical Sciences (U.S.) (Grant GM110535) United States. Defense Advanced Research Projects Agency (Grant 023504-001) 2020-01-21T22:20:04Z 2020-01-21T22:20:04Z 2018-03 2020-01-02T18:58:50Z Article http://purl.org/eprint/type/JournalArticle 1433-7851 https://hdl.handle.net/1721.1/123516 Zhang, Chi et al. "Site-Selective Cysteine-Cyclooctyne Conjugation." Angewandte Chemie International Edition 57, 22 (May 28, 2018): 6459-6463 © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim en http://dx.doi.org/10.1002/anie.201800860 Angewandte Chemie International Edition Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Wiley PMC |
| spellingShingle | Zhang, Chi Dai, Peng Vinogradov, Alexander A. Gates, Zachary P Pentelute, Bradley L. Site-Selective Cysteine-Cyclooctyne Conjugation |
| title | Site-Selective Cysteine-Cyclooctyne Conjugation |
| title_full | Site-Selective Cysteine-Cyclooctyne Conjugation |
| title_fullStr | Site-Selective Cysteine-Cyclooctyne Conjugation |
| title_full_unstemmed | Site-Selective Cysteine-Cyclooctyne Conjugation |
| title_short | Site-Selective Cysteine-Cyclooctyne Conjugation |
| title_sort | site selective cysteine cyclooctyne conjugation |
| url | https://hdl.handle.net/1721.1/123516 |
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