Abstract
Abstract This chapter deals with the use of spectroscopic methods to monitor conformational changes in membrane transporters and kinetic approaches to time-resolve these changes. It outlines the use of fluorescence spectroscopy to monitor ligand-membrane protein interactions and the use of stopped-flow fluorescence spectroscopy to determine mechanisms of ligand binding and translocation. It also outlines the use of rapid-quench techniques for measuring the rate constants governing the translocation of the substrate across the membrane. To a large extent, these studies are illustrated by reference to the kinetics of substrate and inhibitor binding and translocation by various transporters studied in the author’s laboratory.
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