Site‐Selective Cysteine–Cyclooctyne Conjugation |
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| Authors: | Dr. Chi Zhang Dr. Peng Dai Dr. Alexander A. Vinogradov Dr. Zachary P. Gates Prof. Dr. Bradley L. Pentelute |
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| Affiliation: | Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA |
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| Abstract: | 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. |
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| Keywords: | bioconjugation bioorthogonal chemistry cysteine– cyclooctyne reaction dibenzocyclooctyne protein modification |
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