In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation

Nojoumi, Saba; Ma, Ying; Schwagerus, Sergej; Hackenberger, Christian P. R.; Budisa, Nediljko

doi:10.3390/ijms20092299

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In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation

Saba Nojoumi

¹,

Ying Ma

¹,

Sergej Schwagerus

^2,3,

Christian P. R. Hackenberger

^2,3 and

Nediljko Budisa

^1,4,*

Institut für Chemie, Technische Universität Berlin, Müller-Breslau-Str. 10, D-10623 Berlin, Germany

Institut für Chemie der Humboldt-Universität zu Berlin, Brook-Taylor-Str. 2, D-12489 Berlin, Germany

Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Campus Berlin-Buch, Robert-Roessle-Str. 10, D-13125 Berlin, Germany

⁴

Chair of Chemical Synthetic Biology, Department of Chemistry, University of Manitoba, 144 Dysart Rd, Winnipeg, MB R3T 2N2, Canada

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2019, 20(9), 2299; https://doi.org/10.3390/ijms20092299

Received: 18 April 2019 / Revised: 5 May 2019 / Accepted: 7 May 2019 / Published: 9 May 2019

(This article belongs to the Special Issue Expanding and Reprogramming the Genetic Code)

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Abstract

In this study, we report our initial results on in situ biosynthesis of S-allyl-l-homocysteine (Sahc) by simple metabolic conversion of allyl mercaptan in Escherichia coli, which served as the host organism endowed with a direct sulfhydration pathway. The intracellular synthesis we describe in this study is coupled with the direct incorporation of Sahc into proteins in response to methionine codons. Together with O-acetyl-homoserine, allyl mercaptan was added to the growth medium, followed by uptake and intracellular reaction to give Sahc. Our protocol efficiently combined the in vivo synthesis of Sahc via metabolic engineering with reprogrammed translation, without the need for a major change in the protein biosynthesis machinery. Although the system needs further optimisation to achieve greater intracellular Sahc production for complete protein labelling, we demonstrated its functional versatility for photo-induced thiol-ene coupling and the recently developed phosphonamidate conjugation reaction. Importantly, deprotection of Sahc leads to homocysteine-containing proteins—a potentially useful approach for the selective labelling of thiols with high relevance in various medical settings. View Full-Text

Keywords: biorthogonal conjugations; deallylation/deprotection; direct sulfhydration/transsulfuration pathway; homocysteine; methionine metabolism; non-canonical amino acids; O-acetyl-homoserine; reprogrammed translation; S-allyl-homocysteine; selective labelling

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MDPI and ACS Style

Nojoumi, S.; Ma, Y.; Schwagerus, S.; Hackenberger, C.P.R.; Budisa, N. In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation. Int. J. Mol. Sci. 2019, 20, 2299. https://doi.org/10.3390/ijms20092299

AMA Style

Nojoumi S, Ma Y, Schwagerus S, Hackenberger CPR, Budisa N. In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation. International Journal of Molecular Sciences. 2019; 20(9):2299. https://doi.org/10.3390/ijms20092299

Chicago/Turabian Style

Nojoumi, Saba, Ying Ma, Sergej Schwagerus, Christian P.R. Hackenberger, and Nediljko Budisa. 2019. "In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation" International Journal of Molecular Sciences 20, no. 9: 2299. https://doi.org/10.3390/ijms20092299

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In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation

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