Residue choice defines efficiency and influence of bioorthogonal protein modification via genetically encoded strain promoted Click chemistry.
GFP and a FRET compatible dye were used to assess the influence of genetically encoded aryl azide positioning on Click chemistry-based protein conjugation. While modification efficiency of the sampled mutants using a strain promoted reaction varied by as much as ∼10 fold, there was no simple correlation with accessibility of the aryl azide on GFP's surface. One labeled GFP mutant (Gln204AzPhe) exhibited high efficiency FRET (∼90%) and an unprecedented pseudo-Stokes shift of 126 nm.
School of Biosciences, Cardiff University, Cardiff, UK.
Samuel C
SC
Reddington
Eric M
EM
Tippmann
D Dafydd
DD
Jones
D057930:Click Chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D011499:Protein Processing, Post-Translational
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins