Abstract
Chemoselective protein labeling is a valuable tool in the arsenal of modern chemical biology. The unnatural amino acid mutagenesis technology provides a powerful way to site-specifically introduce nonnatural chemical functionalities into recombinant proteins, which can be subsequently functionalized in a chemoselective manner. Even though several strategies currently exist to selectively label recombinant proteins in this manner, there is considerable interest for the development of additional chemoselective reactions that are fast, catalyst-free, use readily available reagents, and are compatible with existing conjugation chemistries. Here we describe a method to express recombinant proteins in E. coli site-specifically incorporating 5-hydroxytryptophan, followed by the chemoselective labeling of this residue using a chemoselective rapid azo-coupling reaction.
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References
Lang K, Chin JW (2014) Bioorthogonal reactions for labeling proteins. ACS Chem Biol 9:16–20. https://doi.org/10.1021/cb4009292
McKay CS, Finn MG (2014) Click chemistry in complex mixtures: bioorthogonal bioconjugation. Chem Biol 21:1075–1101. https://doi.org/10.1016/j.chembiol.2014.09.002
Ramil CP, Lin Q (2013) Bioorthogonal chemistry: strategies and recent developments. Chem Commun (Camb) 49:11007–11022. https://doi.org/10.1039/c3cc44272a
Shih HW, Kamber DN, Prescher JA (2014) Building better bioorthogonal reactions. Curr Opin Chem Biol 21:103–111. https://doi.org/10.1016/j.cbpa.2014.07.002
Sletten EM, Bertozzi CR (2009) Bioorthogonal chemistry: fishing for selectivity in a sea of functionality. Angew Chem Int Ed Engl 48:6974–6998. https://doi.org/10.1002/anie.200900942
Stephanopoulos N, Francis MB (2011) Choosing an effective protein bioconjugation strategy. Nat Chem Biol 7:876–884. https://doi.org/10.1038/nchembio.720
Dumas A, Lercher L, Spicer CD, Davis BG (2015) Designing logical codon reassignment–expanding the chemistry in biology. Chem Sci 6:50–69. https://doi.org/10.1039/C4SC01534G
Kim CH, Axup JY, Schultz PG (2013) Protein conjugation with genetically encoded unnatural amino acids. Curr Opin Chem Biol 17:412–419. https://doi.org/10.1016/j.cbpa.2013.04.017
Lang K, Chin JW (2014) Cellular incorporation of unnatural amino acids and bioorthogonal labeling of proteins. Chem Rev 114:4764–4806. https://doi.org/10.1021/cr400355w
Young DD, Schultz PG (2018) Playing with the molecules of life. ACS Chem Biol 13:854–870. https://doi.org/10.1021/acschembio.7b00974
Italia JS, Zheng Y, Kelemen RE, Erickson SB, Addy PS, Chatterjee A (2017) Expanding the genetic code of mammalian cells. Biochem Soc Trans 45:555–562. https://doi.org/10.1042/BST20160336
Italia JS, Addy PS, Wrobel CJ, Crawford LA, Lajoie MJ, Zheng Y, Chatterjee A (2017) An orthogonalized platform for genetic code expansion in both bacteria and eukaryotes. Nat Chem Biol 13:446–450. https://doi.org/10.1038/nchembio.2312
Addy PS, Erickson SB, Italia JS, Chatterjee A (2017) A chemoselective rapid azo-coupling reaction (CRACR) for unclickable bioconjugation. J Am Chem Soc 139:11670–11673. https://doi.org/10.1021/jacs.7b05125
Addy PS, Italia JS, Chatterjee A (2018) An oxidative bioconjugation strategy targeted to a genetically encoded 5-hydroxytryptophan. Chembiochem 19:1375–1378. https://doi.org/10.1002/cbic.201800111
Italia JS, Addy PS, Erickson SB, Peeler JC, Weerapana E, Chatterjee A (2019) Mutually orthogonal nonsense-suppression systems and conjugation chemistries for precise protein labeling at up to three distinct sites. J Am Chem Soc 141(15):6204–6212
Bruckman MA, Kaur G, Lee LA, Xie F, Sepulveda J, Breitenkamp R, Zhang X, Joralemon M, Russell TP, Emrick T, Wang Q (2008) Surface modification of tobacco mosaic virus with “click” chemistry. Chembiochem 9:519–523. https://doi.org/10.1002/cbic.200700559
Gavrilyuk J, Ban H, Nagano M, Hakamata W, Barbas CF 3rd (2012) Formylbenzene diazonium hexafluorophosphate reagent for tyrosine-selective modification of proteins and the introduction of a bioorthogonal aldehyde. Bioconjug Chem 23:2321–2328. https://doi.org/10.1021/bc300410p
Hooker JM, Kovacs EW, Francis MB (2004) Interior surface modification of bacteriophage MS2. J Am Chem Soc 126:3718–3719. https://doi.org/10.1021/ja031790q
Schlick TL, Ding Z, Kovacs EW, Francis MB (2005) Dual-surface modification of the tobacco mosaic virus. J Am Chem Soc 127:3718–3723. https://doi.org/10.1021/ja046239n
He J, Kimani FW, Jewett JC (2015) A photobasic functional group. J Am Chem Soc 137:9764–9767. https://doi.org/10.1021/jacs.5b04367
Jensen SM, Kimani FW, Jewett JC (2016) Light-activated triazabutadienes for the modification of a viral surface. Chembiochem 17:2216–2219. https://doi.org/10.1002/cbic.201600508
Kimani FW, Jewett JC (2015) Water-soluble triazabutadienes that release diazonium species upon protonation under physiologically relevant conditions. Angew Chem Int Ed Engl 54:4051–4054. https://doi.org/10.1002/anie.201411277
Hutchins BM, Kazane SA, Staflin K, Forsyth JS, Felding-Habermann B, Schultz PG, Smider VV (2011) Site-specific coupling and sterically controlled formation of multimeric antibody fab fragments with unnatural amino acids. J Mol Biol 406:595–603. https://doi.org/10.1016/j.jmb.2011.01.011
Hutchins BM, Kazane SA, Staflin K, Forsyth JS, Felding-Habermann B, Smider VV, Schultz PG (2011) Selective formation of covalent protein heterodimers with an unnatural amino acid. Chem Biol 18:299–303. https://doi.org/10.1016/j.chembiol.2011.01.006
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This work was supported by NIH (R01GM124319 to A.C.).
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Addy, P.S., Erickson, S.B., Italia, J.S., Chatterjee, A. (2019). Labeling Proteins at Site-Specifically Incorporated 5-Hydroxytryptophan Residues Using a Chemoselective Rapid Azo-Coupling Reaction. In: Massa, S., Devoogdt, N. (eds) Bioconjugation. Methods in Molecular Biology, vol 2033. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9654-4_16
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