Abstract
Bioorthogonal chemistry techniques enable the selective and targeted manipulation of living systems. In order to yield universally applicable techniques, it is of great importance for bioorthogonal reactions to take place rapidly, selectively, and with the formation of only benign side products. One of the reactions that match these criteria well is the inverse electron demand Diels-Alder reaction (DAinv) between tetrazines and strained dienophiles. However, even this prime technique comes with the disadvantage of its reactants having limited stability under physiological conditions. In our protocol, an unreactive and therefore stable DAinv diene/dienophile pair reacts rapidly using DNA hybridization as secondary rate-accelerating process. Due to the fluorogenicity of the presented tetrazine rhodamine conjugate, this method enables the selective screening and evaluation of reactant pairs for proximity-mediated bioorthogonal chemistry.
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References
Sletten EM, Bertozzi CR (2009) Bioorthogonal chemistry: fishing for selectivity in a sea of functionality. Angew Chem Int Ed 48(38):6974–6998
Row RD, Prescher JA (2018) Constructing new bioorthogonal reagents and reactions. Acc Chem Res 51(5):1073–1081
Oliveira BL, Guo Z, Bernardes GJL (2017) Inverse electron demand Diels-Alder reactions in chemical biology. Chem Soc Rev 46(16):4895–4950
Blackman ML, Royzen M, Fox JM (2008) Tetrazine ligation: fast bioconjugation based on inverse-electron-demand Diels−Alder reactivity. J Am Chem Soc 130(41):13518–13519
Pipkorn R, Waldeck W, Didinger B, Koch M, Mueller G, Wiessler M, Braun K (2009) Inverse-electron-demand Diels-Alder reaction as a highly efficient chemoselective ligation procedure: Synthesis and function of a BioShuttle for temozolomide transport into prostate cancer cells. J Pept Sci 15(3):235–241
Werther P, Möhler JS, Wombacher R (2017) A bifunctional fluorogenic rhodamine probe for proximity-induced bioorthogonal chemistry. Chem Eur J 23(72):18216–18224
Šečkutė J, Yang J, Devaraj NK (2013) Rapid oligonucleotide-templated fluorogenic tetrazine ligations. Nucleic Acids Res 41(15):e148–e148
Wu H, Alexander SC, Jin S, Devaraj NK (2016) A bioorthogonal near-infrared fluorogenic probe for mRNA detection. J Am Chem Soc 138(36):11429–11432
Anzalone AV, Wang TY, Chen Z, Cornish VW (2013) A common diaryl ether intermediate for the gram-scale synthesis of oxazine and xanthene fluorophores. Angew Chem Int Ed 52(2):650–654
Maiti D, Buchwald SL (2009) Orthogonal Cu-and Pd-based catalyst systems for the O-and N-arylation of aminophenols. J Am Chem Soc 131(47):17423–17429
Eildal JN, Bach A, Dogan J, Ye F, Zhang M, Jemth P, Strømgaard K (2015) Rigidified clicked dimeric ligands for studying the dynamics of the PDZ1-2 supramodule of PSD-95. Chembiochem 16(1):64–69
Wieczorek A, Werther P, Euchner J, Wombacher R (2017) Green-to far-red-emitting fluorogenic tetrazine probes–synthetic access and no-wash protein imaging inside living cells. Chem Sci 8:1506–1510
Yang J, Karver MR, Li W, Sahu S, Devaraj NK (2012) Metal-catalyzed one-pot synthesis of tetrazines directly from aliphatic nitriles and hydrazine. Angew Chem 124(21):5312–5315
Klapars A, Buchwald SL (2002) Copper-catalyzed halogen exchange in aryl halides: an aromatic Finkelstein reaction. J Am Chem Soc 124(50):14844–14845
Acknowledgments
R.W. acknowledges funding from the Deutsche Forschungsgemeinschaft DFG (SPP1623, WO 1888/1–2). We thank Dr. Achim Wieczorek for scientific advice.
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Möhler, J.S., Werther, P., Wombacher, R. (2019). Proximity-Induced Bioorthogonal Chemistry Using Inverse Electron Demand Diels-Alder Reaction. In: Sunbul, M., Jäschke, A. (eds) Proximity Labeling. Methods in Molecular Biology, vol 2008. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9537-0_12
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DOI: https://doi.org/10.1007/978-1-4939-9537-0_12
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