Design, recombinant expression and convenient A-chain N-terminal europium-labelling of a fully active human relaxin-3 analogue.

摘要: Relaxin-3 (also known as INSL7) is a recently identified neuropeptide belonging to the insulin/relaxin superfamily. It plays putative role in regulation of food intake, stress response and reproduction by activating G-protein-coupled receptor, RXFP3. In previous study, we prepared 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)/Eu(3+)-labelled human relaxin-3 tracer for study ligand-receptor interactions, which necessitated complicated site-specific labelling strategy because contains four primary amine moieties, all react with amine-specific modification reagent. To simplify procedure, present created an easily labelled, recombinant analogue only one moiety at A-chain N-terminus. The retained full activity could be labelled various functional probes DOTA/Eu(3+)-labelled high binding affinity its cognate RXFP3, thus represents useful, nonradioactive stable studying interaction RXFP3 natural or synthetic ligands. This new also suitable template design other analogues that can DOTA/Eu(3+) used interactions future.