Labeling of fusion proteins by enzymic incorporation of a coenzyme A derivative into an acyl carrier protein moiety

A method for labeling acyl carrier protein (ACP) fusion proteins using derivs. of CoA is described. The method relies on the transfer of a label from a CoA type substrate to an ACP fusion protein using a holo-acyl carrier protein synthase (ACPS) or a homolog thereof. The method allows detecting and manipulating the fusion protein, both in vitro and in vivo, by attaching mols. to the fusion proteins that introduce a new phys. or chem. property to the fusion protein. Examples of such labels are, among others, spectroscopic probes or reporter mols., affinity tags, mols. generating reactive radicals, cross-linkers, ligands mediating protein-protein interactions or mols. suitable for the immobilization of the fusion protein. Synthesis of a series of reporter mol. conjugates, including digoxigenin, Cy3 and Cy5, with CoA is reported. [on SciFinder (R)]