Site-specific protein labelling and immobilization mediated by microbial transglutaminase.
Microbial transglutaminase (mTG) shows broad substrate specificity that is amenable to in vitro bio-conjugation applications. Herein, test proteins were genetically fused with peptide tags, followed by mTG-mediated propargylation of their reactive Gln residues. The propargylated proteins were subjected to copper-assisted azide-alkyne cycloaddition to demonstrate either fluorescent labelling or immobilization.
Department of Chemistry, University of Ottawa, 10 Marie-Curie, Ottawa, Ontario, Canada K1N 6N5. jkeillor@uottawa.ca.
Samuel K
SK
Oteng-Pabi
Christophe
C
Pardin
Maria
M
Stoica
Jeffrey W
JW
Keillor
D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D002384:Catalysis; D003300:Copper / Q000737:chemistry; D061565:Cycloaddition Reaction; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D005973:Glutamine / Q000737:chemistry* / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry; D010293:Pargyline / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D011437:Propylamines / Q000737:chemistry* / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D013306:Streptomycetaceae / Q000201:enzymology; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
D000480:Alkynes; D001386:Azides; D004800:Enzymes, Immobilized; D009842:Oligopeptides; D011437:Propylamines; D011506:Proteins; D005973:Glutamine; C009054:propargylamine; C037841:tyrosyl-alanyl-glycine; D003300:Copper; D010293:Pargyline; D011503:Transglutaminases