The kinetics of ionophore X-537A-mediated transport of manganese through dipalmitoylphosphatidylcholine vesicles A 1H-NMR study |
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Authors: | Hadassa Degani Sanford Simon Alan C McLaughlin |
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Institution: | 1. Isotope Department, The Weizman Institute of Science, Rehovot Israel;2. Biochemistry Department, SUNY, Stony Brook, NY, U.S.A.;3. Biology Department, Brookhaven National Laboratory, Upton NY 11973 U.S.A. |
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Abstract: | We have studied the kinetics of ionophore X-537A-mediated transport of manganese ions into small unilamellar vesicles formed from dipalmitoylphosphatidylcholine. To follow the transport we used the paramagnetic effect of manganese on the 1H-NMR signal from choline trimethylammonium groups on the inner phospholipid monolayer. The transport of only one manganese ion produces an intravesicular concentration which is high enough (approx. 1 mM) to substantially broaden this signal. The observed signal thus arises predominantly from those vesicles which contain no manganese. Therefore, as manganese is transported into the vesicles the observed signal decreases in intensity, but does not broaden. The initial time-dependence of the intensity of the signal, , can be approximated by the simple first-order rate law: , where is the probability per unit time for the transport of a manganese ion from the external medium to the intravesicular space. From the dependence of on the ionophore X-537A concentration we conclude that manganese is transported into the vesicles via both 1 : 1 and 2 : 1 complexes with ionophore X-537A. At low ratios of ionophore X-537A to vesicles transport via the 1 : 1 complex predominates; at high ratios transport via the 2 : 1 complex predominates. From the dependence of on manganese concentration we determined that under our conditions the equilibration of ionophore X-537A between vesicles is much faster than the transport of manganese through the vesicles. Lastly, from the dependence of on temperature, we conclude that the ionophore X-537A-mediated transport of manganese into the dipalmitoylphosphatidylcholine vesicles is very sensitive to the gel-liquid crystalline phase transition. |
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Keywords: | Dipalmitoylphosphatidylcholine vesicle Ionophore X-537A |
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