The events relating to lanthanide ions enhanced permeability of human erythrocyte membrane: binding, conformational change, phase transition, perforation and ion transport

The binding and uptake of Gd3+ ions by human erythrocytes in vitro were stu died by determining the Gd contents in membrane and in cytosol by means of particle-induced X-ray emission (PIXE) spectrometry. Results obtained from varied incubation time revealed that the Gd3+ ions bind to the membrane pro teins and lipids at first. Gd3+ binding to the membrane lipids and proteins lasts 0 similar to 20 and 20 similar to 100 ms respectively, as shown by t he stopped-flow studies. Then a fraction of Gd3+ ions diffuses through the membrane. The kinetics of Gd3+ binding indicates that the binding to phosph olipids is prior to that to the membrane proteins, but a portion of the lip id-bound Gd3+ redistributed later to the proteins. PIXE studies showed that the entry of Gd3+ increased the influx of Ca2+ and Cl-. By monitoring the changes in fluorescence of proteins and that of the Ln(3+), the uptake of L a3+, Eu3+, Gd3+ and Tb3+ was shown to be a process comprising a series of e vents. Binding to the membrane molecules induces the phase transition of li pid bilayer and conformational changes and aggregation of membrane proteins . Conformational changes of the proteins were characterized by Fourier tran sform IR spectroscopy (FT-IR) deconvolved spectra, i.e. alpha-helix content decreases while beta-sheet increases. ESR spectra of MSL-labeled proteins reflect the aggregation state related with the conformational change. [P-31 ]NMR spectra of membrane lipid bilayer revealed the Ln(3+) ions induced hex agonal (H-II) phase formation. Phase transition and aggregation of membrane proteins cause the formation of domain structure and perforation in the me mbrane. These alterations in membrane structure are responsible for the Ln( 3+) enhanced membrane permeability. Thus the previous Ln(3+) binding will f acilitate the across-membrane transport of other Ln(3+) ions through the me mbrane. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.