Aminopropargyl derivative of terpyridine-bis(methyl-enamine) tetraacetic acid chelate of europium (Eu (TMT)-AP3): a new reagent for fluorescent labelling of proteins and peptides.

The synthesis and photophysical properties of a new terpyridine-based europium(III) chelate (Eu (TMT)-AP3) designed for peptide and protein labelling in aqueous solution phase is described. In order to obtain a stable, easy to handle, versatile and efficient labelling agent, a reactive aminopropargyl arm has been introduced onto the terpyridine moiety. As preliminary biochemical applications the chelate has been 1) efficiently covalently attached onto a representative biomolecule—monoclonal antibody—and 2) converted into iodoacetamido and aldehyde derivatives, and the photoluminescent Eu (TMT)-AP3 was grafted onto cysteine and lysine amino acid residues respectively. These two different solution phase labelling methods yielded original fluorogenic FRET based probes suitable for “in vitro” detection of caspase-3 protease, a key mediator of apoptosis of mammalian cells.