The determination of the kinetic parameters of a carrier mediated transport process in the presence of an unstirred water layer

A procedure is described, based on the Eadie-Hofstee plot, from which it is possible to determine the Michaelis constant and the maximum velocity of a membrane bound transport process that is separated from the substrate in the bulk solution by an unstirred layer. This can be done without knowing the magnitude of the latter provided that its effective thickness can be varied by altering the rate of stirring. Further, if the affinity of the carrier for the substrate is sufficiently large, then, it is possible to determine the rate constant and the effective concentration of the membrane bound carrier.