Survey of carrier methodology: Strategy for identification, isolation, and characterization of transport systems

  • Klingenberg M
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In this chapter, methods are examined using three different transport systems: glucose transport in erythrocytes, ADP/ATP exchange in mitochondria, and lactose transport in Escherichia coli. For determining the transport function of a carrier and the dependence on environmental factors it is useful to isolate the protein and to study its function after reincorporation into artificial phospholipid vesicles. Another reason for isolating membrane protein is to characterize its molecular properties and eventually its molecular mechanism. The isolation of a specific membrane protein requires first its identification among the other membrane proteins. The identification of biomembrane carriers serves to characterize the transport processes, their biological implications, and their mechanism. Together they form the groundwork for kinetic studies, either in the original intact membrane or in the reconstituted system. These include functional studies on the role of activators, inhibitors, and regulators, the influence of membrane potential, the coordination with other transport processes, and the mechanism of transport as described in kinetic terms. © 1989, Elsevier Inc. All rights reserved.

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