Biophysical Journal
Volume 83, Issue 4, October 2002, Pages 2118-2125
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Probing Red Cell Membrane Cholesterol Movement with Cyclodextrin

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Abstract

We probed the kinetics with which cholesterol moves across the human red cell bilayer and exits the membrane using methyl-β-cyclodextrin as an acceptor. The fractional rate of cholesterol transfer (% s−1) was unprecedented, the half-time at 37°C being ∼1 s. The kinetics observed under typical conditions were independent of donor concentration and directly proportional to acceptor concentration. The rate of exit of membrane cholesterol fell hyperbolically to zero with increasing dilution. The energy of activation for cholesterol transfer was the same at high and low dilution; namely, 27–28 Kcal/mol. This behavior is not consistent with an exit pathway involving desorption followed by aqueous diffusion to acceptors nor with a simple one-step collision mechanism. Rather, it is that predicted for an activation–collision mechanism in which the reversible partial projection of cholesterol molecules out of the bilayer precedes their collisional capture by cyclodextrin. Because the entire membrane pool was transferred in a single first-order process under all conditions, we infer that the transbilayer diffusion (flip-flop) of cholesterol must have proceeded faster than its exit, i.e., with a half-time of <1 s at 37°C.

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