A pyrrolysine analogue for protein click chemistry.
Ignoring the STOP sign: A pyrrolysine analogue bearing a terminal alkyne was site-specifically incorporated into recombinant calmodulin (CaM) through a UAG codon. The resulting protein was labeled with an azide-containing dye using a copper(I)-catalyzed click reaction. Subsequent application of an orthogonal cysteine tagging method yielded a CaM labeled with two distinct fluorophores that enabled its study by FRET spectroscopy.
Fekner
Tomasz
T
Department of Chemistry, The Ohio State University, 484 West 12th Avenue, Columbus, OH 43210, USA.
Li
Xin
X
Lee
Marianne M
MM
Chan
Michael K
MK
D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000138:chemical synthesis / Q000737:chemistry; D002147:Calmodulin / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D002384:Catalysis; D018388:Codon, Terminator / Q000235:genetics; D003300:Copper / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D008239:Lysine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry*
D000480:Alkynes; D001386:Azides; D002147:Calmodulin; D018388:Codon, Terminator; D011994:Recombinant Proteins; D003300:Copper; C456839:pyrrolysine; D008239:Lysine