A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling

Bioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an...

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Autores principales:	Birthe Meineke, Johannes Heimgärtner, Alexander J. Craig, Michael Landreh, Lindon W. K. Moodie, Simon J. Elsässer
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	genetic code expansion amber suppression noncanonical amino acid bioorthogonal chemistry click chemistry copper catalyzed azide–alkyne cycloaddition (CuAAC) Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/80dfbc6fdcfb4fc3aa01db18bc5f6838
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spelling	oai:doaj.org-article:80dfbc6fdcfb4fc3aa01db18bc5f68382021-11-11T10:21:53ZA Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling2296-264610.3389/fchem.2021.768535https://doaj.org/article/80dfbc6fdcfb4fc3aa01db18bc5f68382021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fchem.2021.768535/fullhttps://doaj.org/toc/2296-2646Bioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an efficient method for incorporating such chemical handles into proteins on the ribosome, in which noncanonical amino acids (ncAAs) are site specifically introduced into the polypeptide in response to an amber (UAG) stop codon. A variety of ncAA structures containing azides or alkynes have been proven useful for performing CuAAC chemistry on proteins. To improve CuAAC efficiency, biologically incorporated alkyne groups can be reacted with azide substrates that contain copper-chelating groups. However, the direct incorporation of copper-chelating azides into proteins has not been explored. To remedy this, we prepared the ncAA paz-lysine (PazK), which contains a picolyl azide motif. We show that PazK is efficiently incorporated into proteins by amber suppression in mammalian cells. Furthermore, PazK-labeled proteins show improved reactivity with alkyne reagents in CuAAC.Birthe MeinekeBirthe MeinekeJohannes HeimgärtnerJohannes HeimgärtnerAlexander J. CraigMichael LandrehLindon W. K. MoodieLindon W. K. MoodieSimon J. ElsässerSimon J. ElsässerFrontiers Media S.A.articlegenetic code expansionamber suppressionnoncanonical amino acidbioorthogonal chemistryclick chemistrycopper catalyzed azide–alkyne cycloaddition (CuAAC)ChemistryQD1-999ENFrontiers in Chemistry, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	genetic code expansion amber suppression noncanonical amino acid bioorthogonal chemistry click chemistry copper catalyzed azide–alkyne cycloaddition (CuAAC) Chemistry QD1-999
spellingShingle	genetic code expansion amber suppression noncanonical amino acid bioorthogonal chemistry click chemistry copper catalyzed azide–alkyne cycloaddition (CuAAC) Chemistry QD1-999 Birthe Meineke Birthe Meineke Johannes Heimgärtner Johannes Heimgärtner Alexander J. Craig Michael Landreh Lindon W. K. Moodie Lindon W. K. Moodie Simon J. Elsässer Simon J. Elsässer A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
description	Bioorthogonal chemistry allows rapid and highly selective reactivity in biological environments. The copper-catalyzed azide–alkyne cycloaddition (CuAAC) is a classic bioorthogonal reaction routinely used to modify azides or alkynes that have been introduced into biomolecules. Amber suppression is an efficient method for incorporating such chemical handles into proteins on the ribosome, in which noncanonical amino acids (ncAAs) are site specifically introduced into the polypeptide in response to an amber (UAG) stop codon. A variety of ncAA structures containing azides or alkynes have been proven useful for performing CuAAC chemistry on proteins. To improve CuAAC efficiency, biologically incorporated alkyne groups can be reacted with azide substrates that contain copper-chelating groups. However, the direct incorporation of copper-chelating azides into proteins has not been explored. To remedy this, we prepared the ncAA paz-lysine (PazK), which contains a picolyl azide motif. We show that PazK is efficiently incorporated into proteins by amber suppression in mammalian cells. Furthermore, PazK-labeled proteins show improved reactivity with alkyne reagents in CuAAC.
format	article
author	Birthe Meineke Birthe Meineke Johannes Heimgärtner Johannes Heimgärtner Alexander J. Craig Michael Landreh Lindon W. K. Moodie Lindon W. K. Moodie Simon J. Elsässer Simon J. Elsässer
author_facet	Birthe Meineke Birthe Meineke Johannes Heimgärtner Johannes Heimgärtner Alexander J. Craig Michael Landreh Lindon W. K. Moodie Lindon W. K. Moodie Simon J. Elsässer Simon J. Elsässer
author_sort	Birthe Meineke
title	A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
title_short	A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
title_full	A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
title_fullStr	A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
title_full_unstemmed	A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling
title_sort	genetically encoded picolyl azide for improved live cell copper click labeling
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/80dfbc6fdcfb4fc3aa01db18bc5f6838
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A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling

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