Phosphorus-31 and carbon-13 nuclear magnetic resonance studies of divalent cation binding to phosphatidylserine membranes: use of cobalt as a paramagnetic probe.
The paramagnetic divalent cation cobalt has large and well-understood effects on NMR signals from ligands bound in the first coordination sphere, i.e., inner-sphere ligands, and we have used these effects to identify divalent cation binding sites at the surface of phosphatidylserine membranes. 31P NMR results show that 13% of the bound cobalt ions are involved in inner-sphere complexes with the phosphodiester group, while 13C NMR results show that 54% of the bound cobalt ions are involved in unidentate inner sphere complexes with the carboxyl group. No evidence is found for cobalt binding to the carbonyl groups, but proton release studies suggest that 32% of the bound cobalt ions are involved in chelate complexes that contain both the carboxyl and the amine groups. All (i.e., 13% + 54% + 32% = 99%) of the bound cobalt ions can thus be accounted for in terms of inner sphere complexes with the phosphodiester group or the carboxyl group. We suggest that the unidentate inner-sphere complex between cobalt and the carboxyl group of phosphatidylserine and the inner-sphere complex between cobalt and the phosphodiester group of phosphatidylserine provide reasonable models for complexes between alkaline earth cations and phosphatidylserine membranes.
D000818:Animals; D001665:Binding Sites; D001921:Brain; D002247:Carbon Isotopes; D002417:Cattle; D003035:Cobalt; D008081:Liposomes; D009682:Magnetic Resonance Spectroscopy; D010718:Phosphatidylserines; D010761:Phosphorus Radioisotopes
D002247:Carbon Isotopes; D008081:Liposomes; D010718:Phosphatidylserines; D010761:Phosphorus Radioisotopes; D003035:Cobalt