Catalyst-free and site-specific one-pot dual-labeling of a protein directed by two genetically incorporated noncanonical amino acids.
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abstract
The genetic incorporation of one azide-containing and one keto-containing noncanonical amino acids into a protein at one amber and one ochre mutation sites respectively followed by their orthogonal reactions with one hydroxylamine-containing and one cyclooctyne-containing dyes allows highly efficient one-pot site-specific dual labeling of the protein in a catalyst-free fashion.