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. 2003 May;121(5):413–425. doi: 10.1085/jgp.200308803

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Copyright © 2003, The Rockefeller University Press

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Figure 5. — Water-permeability reductions induced by annexin A4 binding are due to rigidification of the outer leaflet of the liposomal membrane. Polarized fluorescence anisotropy was performed on PC:PS liposomes incubated for 30 min with 2 μM NBD-HPC. Polarized excitation and emission fluorescence was monitored at 465 and 530 nm, respectively. Addition of 1 mM Ca²⁺ or 15 μg Anx4 alone for 10 min did not alter fluorescence anisotropy. Addition of Ca²⁺ and Anx4 (C/A) simultaneously resulted in a significant rigidification (as measured by increased anisotropy) of the outer leaflet of the liposomal membrane. Subsequent addition of EDTA resulted in an increase in fluidity to values exhibited by the control. (n = 3, data shown is mean ± SEM; ***, P < 0.001).