Rapid determination of the transbilayer distribution of NBD-phospholipids in erythrocyte membranes with dithionite.

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.

journal_name

Mol Membr Biol

authors

Pomorski T,Herrmann A,Zachowski A,Devaux PF,Müller P

doi

10.3109/09687689409161028

subject

Has Abstract

pub_date

1994-01-01 00:00:00

pages

39-44

issue

1

eissn

0968-7688

issn

1464-5203

journal_volume

11

pub_type

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