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  • Title: Rapid determination of the transbilayer distribution of NBD-phospholipids in erythrocyte membranes with dithionite.
    Author: Pomorski T, Herrmann A, Zachowski A, Devaux PF, Müller P.
    Journal: Mol Membr Biol; 1994; 11(1):39-44. PubMed ID: 8019600.
    Abstract:
    The assessment of the transverse distribution and mobility of NBD-labelled phospholipid analogues in biological membranes by selective chemical destruction of fluorescent label in the outer monolayer with dithionite has been investigated using resealed erythrocyte ghosts as a model system. The distribution of those analogues can be determined in < 30 s directly in the cell suspension provided the permeation of dithionite across the membrane is suppressed. The results were compared with data on translocation of either NBD- or spin-labelled phospholipid analogues obtained with the technique of back exchange to BSA. It is shown that the passage of dithionite can be mediated by anion-transport systems such as band 3 which is inhibited by DIDS. Appropriate conditions for the applicability of the assay were elucidated also using resealed ghosts having fluorescent NBD-taurine in the intracellular lumen. The application of the assay to measure fast translocation processes, e.g. those mediated by the aminophospholipid translocase, is described.
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