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

• Published in: Organic & biomolecular chemistry Vol. 4; no. 22; pp. 4165 - 4177
• Main Authors: Poupart, Severine, Boudou, Cedric, Peixoto, Philippe, Massonneau, Marc, Renard, Pierre-Yves, Romieu, Anthony
• Format: Journal Article
• Language: English
• Published: CAMBRIDGE Royal Soc Chemistry 01.01.2006
• Subjects: Affinity Labels - chemical synthesis; Affinity Labels - chemistry; Chelating Agents - chemical synthesis; Chelating Agents - chemistry; Chemistry; Chemistry, Organic; Europium - chemistry; Fluorescence Resonance Energy Transfer; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Ligands; Molecular Structure; Organometallic Compounds - chemical synthesis; Organometallic Compounds - chemistry; Peptides - chemistry; Photochemistry; Physical Sciences; Proteins - chemistry; Pyridines - chemistry; Science & Technology; Stereoisomerism; Time Factors; APOPTOSIS; TIME-RESOLVED FLUORESCENCE; LUMINESCENT EUROPIUM(III); SOLID-PHASE SYNTHESIS; CASPASE-3; CROSS-LINKING; LANTHANIDE COMPLEXES; INHIBITORS; AQUEOUS-SOLUTION; PROTEASE ASSAYS
• ISSN: 1477-0520; 1477-0539
• DOI: 10.1039/b612805j

The synthesis and photophysical properties of a new terpyridine-based europium(III) chelate (Eu (TMT)-AP(3)) 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)-AP(3) 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.