title: Competition between Li+ and Mg2+ for the phosphate groups in the human erythrocyte membrane and ATP: an NMR and fluorescence study.
abstract: We investigated the mechanism of competition between Li+ and Mg2+ in Li(+)-loaded human red blood cells (RBCs) by making 7Li and 31P NMR and fluorescence measurements. We used 7Li NMR relaxation times to probe Li+ binding to the human RBC membrane and ATP; an increase in Mg2+ concentration caused an increase in both 7Li T1 and T2 values in packed Li(+)-loaded RBCs, in suspensions of Li(+)-loaded RBC ghosts, in suspensions of Li(+)-containing RBC membrane, and in aqueous solutions of ATP, indicating competition between Li+ and Mg2+ for binding sites in the membrane and ATP. We found that increasing concentrations of either Li+ or Mg2+ in the presence of human RBC membrane caused an increase in the 31P NMR chemical shift anisotropy parameter, which describes the observed axially symmetric powder pattern, indicating metal ion binding to the phosphate groups in the membrane. Competition between Li+ and Mg2+ for phosphate groups in ATP and in the RBC membrane was also observed by both fluorescence measurements and 31P NMR spectroscopy at low temperature. The ratio of the stoichiometric binding constants of Mg2+ to Li+ to the RBC membrane was approximately 20; the ratio of the conditional binding constants in the presence of a free intracellular ATP concentration of 0.2 mM was approximately 4, indicating that Li+ competes for approximately 20% of the Mg(2+)-binding sites in the RBC membrane. Our results indicate that, regardless of the spectroscopic method used, Li+ competes with Mg2+ for phosphate groups in both ATP and the RBC membrane; the extent of metal ion competition for the phosphate head groups of the phospholipids in the RBC membrane is enhanced by the presence of ATP. Competition between Li+ and Mg2+ for anionic phospholipids or Mg(2+)-activated proteins present in cell membranes may constitute the basis of a general molecular mechanism for Li+ action in human tissues.
authors.affiliation: Department of Chemistry, Loyola University of Chicago, Illinois 60626.
authors.forename: D
authors.initials: D
authors.lastname: Mota de Freitas
authors.forename: L
authors.initials: L
authors.lastname: Amari
authors.forename: C
authors.initials: C
authors.lastname: Srinivasan
authors.forename: Q
authors.initials: Q
authors.lastname: Rong
authors.forename: R
authors.initials: R
authors.lastname: Ramasamy
authors.forename: A
authors.initials: A
authors.lastname: Abraha
authors.forename: C F
authors.initials: CF
authors.lastname: Geraldes
authors.forename: M K
authors.initials: MK
authors.lastname: Boyd
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D001667:Binding, Competitive; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005454:Fluorescence Polarization; D006801:Humans; D066298:In Vitro Techniques; D008094:Lithium / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D010710:Phosphates / Q000378:metabolism*
chemical_list: D010710:Phosphates; D000255:Adenosine Triphosphate; D008094:Lithium; D008274:Magnesium