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

Meineke, BirtheKarolinska Institutet,Karolinska Inst, Dept Med Biochem & Biophys, Div Genome Biol, Sci Life Lab, Stockholm, Sweden.;Karolinska Inst, Ming Wai Lau Ctr Reparat Med, Stockholm Node, Stockholm, Sweden. (författare)

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.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
genetic code expansion
amber suppression
noncanonical amino acid
bioorthogonal chemistry
click chemistry
copper catalyzed azide-alkyne cycloaddition (CuAAC)

Heimgärtner, JohannesKarolinska Inst, Dept Med Biochem & Biophys, Div Genome Biol, Sci Life Lab, Stockholm, Sweden.;Karolinska Inst, Ming Wai Lau Ctr Reparat Med, Stockholm Node, Stockholm, Sweden. (författare)
Craig, Alexander J.Uppsala universitet,Läkemedelsdesign och läkemedelsutveckling(Swepub:uu)alecr417 (författare)
Landreh, MichaelKarolinska Institutet,Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Sci Life Lab, Stockholm, Sweden. (författare)
Moodie, Lindon W. K.Uppsala universitet,Läkemedelsdesign och läkemedelsutveckling,Uppsala antibiotikacentrum(Swepub:uu)linmo351 (författare)
Elsässer, Simon J.Karolinska Institutet,Karolinska Inst, Dept Med Biochem & Biophys, Div Genome Biol, Sci Life Lab, Stockholm, Sweden.;Karolinska Inst, Ming Wai Lau Ctr Reparat Med, Stockholm Node, Stockholm, Sweden. (författare)
Karolinska InstitutetKarolinska Inst, Dept Med Biochem & Biophys, Div Genome Biol, Sci Life Lab, Stockholm, Sweden.;Karolinska Inst, Ming Wai Lau Ctr Reparat Med, Stockholm Node, Stockholm, Sweden. (creator_code:org_t)

Av författaren/redakt...: Meineke, Birthe; Heimgärtner, Joh ...; Craig, Alexander ...; Landreh, Michael; Moodie, Lindon W ...; Elsässer, Simon ...

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