title: The events relating to lanthanide ions enhanced permeability of human erythrocyte membrane: binding, conformational change, phase transition, perforation and ion transport.
abstract: The binding and uptake of Gd3+ ions by human erythrocytes in vitro were studied 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 proteins and lipids at first. Gd3+ binding to the membrane lipids and proteins lasts 0 approximately 20 and 20 approximately 100 ms respectively, as shown by the stopped-flow studies. Then a fraction of Gd3+ ions diffuses through the membrane. The kinetics of Gd3+ binding indicates that the binding to phospholipids is prior to that to the membrane proteins, but a portion of the lipid-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 Ln3+, the uptake of La3+, Eu3+, Gd3+ and Tb3+ was shown to be a process comprising a series of events. Binding to the membrane molecules induces the phase transition of lipid bilayer and conformational changes and aggregation of membrane proteins. Conformational changes of the proteins were characterized by Fourier transform 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. [31P]NMR spectra of membrane lipid bilayer revealed the Ln3+ ions induced hexagonal (H(II)) phase formation. Phase transition and aggregation of membrane proteins cause the formation of domain structure and perforation in the membrane. These alterations in membrane structure are responsible for the Ln3+ enhanced membrane permeability. Thus the previous Ln3+ binding will facilitate the across-membrane transport of other Ln3+ ions through the membrane.
authors.affiliation: National Research Laboratories of Natural and Biomimetic Drugs, Beijing Medical University, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Cheng
authors.forename: H
authors.initials: H
authors.lastname: Yao
authors.forename: H
authors.initials: H
authors.lastname: Lin
authors.forename: J
authors.initials: J
authors.lastname: Lu
authors.forename: R
authors.initials: R
authors.lastname: Li
authors.forename: K
authors.initials: K
authors.lastname: Wang
mesh_terms: D001798:Blood Proteins / Q000378:metabolism; D002463:Cell Membrane Permeability; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D017136:Ion Transport; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008674:Metals, Rare Earth / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D001798:Blood Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D008674:Metals, Rare Earth