29/05/2019 - 12:00 - 11:00
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2019-05-29 11:00:00
2019-05-29 12:00:00
Seminar: Genetic code expansion technology for biochemical studies
S E M I N A R
Wednesday 29/05/19, 11:00 am
Building 211, seminar room
SPEAKER:
Dr. Eyal Arbely
Department of Chemistry and the National Institute for Biotechnology
Ben-Gurion University of the Negev
TOPIC:
Genetic code expansion technology for biochemical studies
The chemical repertoire of genetically encoded amino acids can now be expanded beyond the 20 canonical amino acids using genetic code expansion (GCE) technology. By utilizing an orthogonal aminoacyl-tRNA synthetase/tRNA pair and an alternative codon (e.g., the UAG stop codon) GCE enables the site-specific incorporation of NCAAs with unique chemical groups into ribosomally synthesized proteins. One of the emerging applications of GCE is the incorporation of NCAAs bearing functional groups at a single site in the protein for subsequent chemoselective reactions. We used an orthogonal and evolved pyrrolysine tRNA synthetase/tRNACUA pair to genetically encode the incorporation of strained alkenes and alkynes into proteins expressed in cultured mammalian cells. These proteins were then labeled with tetrazine-conjugated fluorescent organic dyes via an inverse electron demand Diels-Alder reaction. The site-specific and biorthogonal fluoregenic reaction enabled fluorescent imaging of a-tubulin and membrane anchored proteins at high resolution in live cells, and provided a superior alternative for fluorescence imaging based on fluorescent proteins. Another attractive application of GCE is the incorporation of post-translationally modified amino acids that allows the synthesis of homogenously and site-specifically modified proteins in bacteria and cultured mammalian cells. In particular, we genetically encoded the incorporation of Ne-acetyl lysine and its derivatives, in order to study the structural and functional role of lysine acetylation in the regulation of transcription.
Abstract
Department of Chemistry
chemistry.office@biu.ac.il
Asia/Jerusalem
public
S E M I N A R
Wednesday 29/05/19, 11:00 am
Building 211, seminar room
SPEAKER:
Dr. Eyal Arbely
Department of Chemistry and the National Institute for Biotechnology
Ben-Gurion University of the Negev
TOPIC:
Genetic code expansion technology for biochemical studies
The chemical repertoire of genetically encoded amino acids can now be expanded beyond the 20 canonical amino acids using genetic code expansion (GCE) technology. By utilizing an orthogonal aminoacyl-tRNA synthetase/tRNA pair and an alternative codon (e.g., the UAG stop codon) GCE enables the site-specific incorporation of NCAAs with unique chemical groups into ribosomally synthesized proteins. One of the emerging applications of GCE is the incorporation of NCAAs bearing functional groups at a single site in the protein for subsequent chemoselective reactions. We used an orthogonal and evolved pyrrolysine tRNA synthetase/tRNACUA pair to genetically encode the incorporation of strained alkenes and alkynes into proteins expressed in cultured mammalian cells. These proteins were then labeled with tetrazine-conjugated fluorescent organic dyes via an inverse electron demand Diels-Alder reaction. The site-specific and biorthogonal fluoregenic reaction enabled fluorescent imaging of a-tubulin and membrane anchored proteins at high resolution in live cells, and provided a superior alternative for fluorescence imaging based on fluorescent proteins. Another attractive application of GCE is the incorporation of post-translationally modified amino acids that allows the synthesis of homogenously and site-specifically modified proteins in bacteria and cultured mammalian cells. In particular, we genetically encoded the incorporation of Ne-acetyl lysine and its derivatives, in order to study the structural and functional role of lysine acetylation in the regulation of transcription.
Abstract
