Abstract: | We have characterized the quenching of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine by Co2+ in egg phosphatidylcholine (PC) lipid bilayer vesicles. The quenching constant obtained is 59 M-1. We demonstrate one use of this fluorescence quenching technique by measuring intrinsic and X537A-mediated transmembrane Co2+ fluxes in large unilamellar PC vesicles. The intrinsic rate constant for Co2+ flux we measure is 3 X 10(-6) S-1. We confirm that the neutral Co approximately (X537A)2 complex is the main component of the X537A-mediated cobalt flux. Since this method measures the concentration of Co2+ at the site of the fluorophore, it is generally applicable to the measurement of paramagnetic ion concentrations in the region of the membrane/water interface. |