Site-specific sortase-catalyzed lipidation of proteins

A methodology for the enzymatic ligation of proteins with a lipid is proposed, a feature of which is the use of a hydrophilized [thanks to a CMG(2) spacer] and therefore highly water-soluble lipid. Its H₂N-Gly₅ terminus and the recombinant protein amino acid sequence LPETG are substrates of the enzyme sortase A. Two lipid-ligated proteins, red fluorescent mCherry and Protein A, are shown to be inserted into the cell membrane.