Interactions of the hormone oxytocin with divalent metal ions.

The interaction of the cyclic nonapeptide oxytocin (OT) with a number of alkaline earth and divalent transition metal ions (X(2+)) was examined employing mass spectrometry (MS) and ion mobility spectrometry (IMS) techniques in combination with molecular dynamics (MD) and density functional theory (DFT) calculations. Under acidic conditions it was found that OT exhibits an exceptionally strong affinity for all divalent metal ions resulting in strong [OT + X](2+) peaks in the mass spectrum. Under basic conditions only Cu(2+) and Ni(2+)-OT complexes were detected and these were singly, doubly, triply, or quadruply deprotonated. Collision-induced dissociation of the [OT - 3H + Cu](-) complex yielded exclusively C-terminal Cu(2+)-containing fragments (Cu(2+)fragment(3-)), suggesting that the Cu(2+) ligation site includes deprotonated C-terminal backbone amide nitrogen atoms and the N-terminal amino nitrogen atom in [OT - 3H + Cu](-). MD and DFT calculations indicate a square-planar complex is consistent with these observations and with experimental collision cross sections. MD and DFT calculations also indicate either an octahedral or trigonal-bipyramidal complex between Zn(2+) and OT is lowest in energy with carbonyl oxygens being the primary ligation sites. Both complexes yield cross sections in agreement with experiment. The biological impact of the structural changes induced in OT by divalent metal ion coodination is discussed.