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Pumps, Channels, and Transporters: Methods of Functional Analysis
Zitatschlüssel TF2015_1
Autor Thomas Friedrich
Seiten 89-120
Jahr 2015
DOI 10.1002/9781119085126.ch4
Herausgeber Ronald J. Clarke and Mohammed A.A. Khalid
Verlag John Wiley & sons, Inc.
Kapitel Probing Conformational Transitions of Membrane Proteins with Voltage Clamp Fluoremetry (VCF)
Zusammenfassung This chapter describes the technique of voltage clamp fluorometry (VCF), which has been widely applied to investigate conformational transitions in, for example, voltage-gated cation channels and electrogenic ion transporters. VCF expands the electrophysiological toolbox to investigate molecular processes in membrane proteins that are not directly or indirectly connected to charge translocation across the membrane. Conformational transitions, either induced by biochemical stimulation or by voltage pulses during two-electrode voltage clamping, entail changes in the microenvironment of the attached fluorophore that eventually result in changes in the fluorescence intensity, reflecting the evolution of conformational changes in time. The typical components of a VCF recording system are depicted in the chapter. Members of the large superfamily of P-type ATPases are found in all the kingdoms of life and share distinct structural and functional features. The technique is particularly advantageous for the study of membrane transporters with net electroneutral activity.
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