A simple experimental approach to the determination of carrier transport parameters for unlabeled substrate analogs.

A method is described by which affinities and transport rates for unlabeled substrate analogs are readily determined, and which is based on the effect of an unlabeled analog upon the rate of transport of a labeled substrate present at a low concentration on the trans side of the membrane. The procedure is widely applicable since it does not depend on assumptions about rrate-limiting steps and holds for both active and non-active systems. Here it is applied in an experimental study of the facilitated diffusion system for choline in erythrocytes, and it is shown that the transport parameters for a test substrate obtained by this method are the same as those found when the transport of the substrate is followed directly.