Page 136 - Tyrosine-Based Bioconjugations - Jorick Bruins

Appendix
It is possible that the installed tyrosine tag might be poorly accessible for mushroom tyrosinase, it is then advised to elongate the tag with an additional G4S-spacer sequence preceding the G4Y-tag, effectively yielding a G4SG4Y-tag. This can be repeated with several G4S units to further increase the spacing and improve the steric accessibility. This can lead to increased reaction rates and improved overall conjugation efficiencies.
14. Depending on the size of the protein a different amount of protein may be used for denaturation, as well as a different acrylamide concentration and protein ladder. Depending on the protein stability, different incubation times may be applied. The same holds true for DTT denaturation and subsequent HPLC analysis. Different denaturation times and different columns might be necessary.
References
(1) Borrmann, A., Fatunsin, O., Dommerholt, J., Jonker, A. M., Lowik, D. W. P. M., van Hest, J. C. M., and van Delft, F. L., Strain-promoted oxidation-controlled cyclooctyne-1,2-quinone cycloaddition (SPOCQ) for fast and activatable protein conjugation. Bioconjugate Chem. 2015, 26 (2), 257-61.
(2) Bruins, J. J., Westphal, A. H., Albada, B., Wagner, K., Bartels, L., Spits, H., van Berkel, W. J. H., and van Delft, F. L., Inducible, Site-Specific Protein Labeling by Tyrosine Oxidation- Strain-Promoted (4+2) Cycloaddition. Bioconjugate Chem. 2017, 28 (4), 1189-93.
(3) Bruins, J. J., Blanco-Ania, D., van der Doef, V., van Delft, F. L., and Albada, B., Orthogonal, dual protein labelling by tandem cycloaddition of strained alkenes and alkynes to ortho- quinones and azides. Chem. Commun. 2018, 54 (53), 7338-41.
(4) Dommerholt, J., Schmidt, S., Temming, R., Hendriks, L. J. A., Rutjes, F. P. J. T., van Hest, J. C. M., Lefeber, D. J., Friedl, P., and van Delft, F. L., Readily accessible bicyclononynes for bioorthogonal labeling and three-dimensional imaging of living cells. Angew. Chem. Int. Ed. 2010, 49 (49), 9422-5.
(5) Taylor, M. T., Blackman, M. L., Dmitrenko, O., and Fox, J. M., Design and Synthesis of Highly Reactive Dienophiles for the Tetrazine trans-Cyclooctene Ligation. J. Am. Chem. Soc. 2011, 133 (25), 9646-9.
(6) Kim, S., Sung, B. H., Kim, S. C., and Lee, H. S., Genetic incorporation of l- dihydroxyphenylalanine (DOPA) biosynthesized by a tyrosine phenol-lyase. Chem. Commun. 2018, 54 (24), 3002-5.
(7) Minamihata, K., Goto, M., and Kamiya, N., Site-specific protein cross-linking by peroxidase- catalyzed activation of a tyrosine-containing peptide tag. Bioconjugate Chem. 2011, 22 (1), 74-81.
(8) Long, M. J. C., and Hedstrom, L., Mushroom tyrosinase oxidizes tyrosine-rich sequences to allow selective protein functionalization. ChemBioChem 2012, 13 (12), 1818-25.
(9) Bruins, J. J., Albada, B., and van Delft, F., ortho-Quinones and Analogues Thereof: Highly Reactive Intermediates for Fast and Selective Biofunctionalization. Chem. - Eur. J. 2018, 24 (19), 4749-56.
(10) Debets, M. F., Van Berkel, S. S., Dommerholt, J., Dirks, A. J., Rutjes, F. P. J. T., and Van Delft, F. L., Bioconjugation with Strained Alkenes and Alkynes. Acc. Chem. Res. 2011, 44 (9), 805- 15.
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