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2019-05-09Zeitschriftenartikel DOI: 10.18452/25467
In-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translation
dc.contributor.authorNojoumi, Saba
dc.contributor.authorMa, Ying
dc.contributor.authorSchwagerus, Sergej
dc.contributor.authorHackenberger, Christian
dc.contributor.authorBudisa, Nediljko
dc.date.accessioned2022-11-11T14:18:47Z
dc.date.available2022-11-11T14:18:47Z
dc.date.issued2019-05-09none
dc.date.updated2022-09-07T06:57:16Z
dc.identifier.urihttp://edoc.hu-berlin.de/18452/26153
dc.description.abstractIn 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.eng
dc.language.isoengnone
dc.publisherHumboldt-Universität zu Berlin
dc.rights(CC BY 4.0) Attribution 4.0 Internationalger
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectbiorthogonal conjugationseng
dc.subjectdeallylation/deprotectioneng
dc.subjectdirect sulfhydration/transsulfuration pathwayeng
dc.subjecthomocysteineeng
dc.subjectmethionine metabolismeng
dc.subjectnon-canonical amino acidseng
dc.subjectO-acetyl-homoserineeng
dc.subjectreprogrammed translationeng
dc.subjectS-allyl-homocysteineeng
dc.subjectselective labellingeng
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftennone
dc.subject.ddc570 Biologienone
dc.titleIn-Cell Synthesis of Bioorthogonal Alkene Tag S-Allyl-Homocysteine and Its Coupling with Reprogrammed Translationnone
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-110-18452/26153-5
dc.identifier.doihttp://dx.doi.org/10.18452/25467
dc.type.versionpublishedVersionnone
local.edoc.pages20none
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
dc.description.versionPeer Reviewednone
dc.identifier.eissn1422-0067
dcterms.bibliographicCitation.doi10.3390/ijms20092299none
dcterms.bibliographicCitation.journaltitleInternational journal of molecular sciencesnone
dcterms.bibliographicCitation.volume20none
dcterms.bibliographicCitation.issue9none
dcterms.bibliographicCitation.articlenumber2299none
dcterms.bibliographicCitation.originalpublishernameMolecular Diversity Preservation Internationalnone
dcterms.bibliographicCitation.originalpublisherplaceBaselnone
bua.departmentMathematisch-Naturwissenschaftliche Fakultätnone

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