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Introduction to Solid Supported Membrane Based Electrophysiology
Published on: May 11, 2013
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SSM-Based Electrophysiology for Transporter Research.
Andre Bazzone1, Maria Barthmes2, Klaus Fendler3
1Max Planck Institute of Biophysics, Frankfurt/Main, Germany; Nanion Technologies GmbH, Munich, Germany.
Methods in Enzymology
|August 7, 2017
Summary
Solid-supported membrane (SSM)-based electrophysiology offers a sensitive, cell-free method for characterizing electrogenic membrane proteins. This technique enables detailed kinetic analysis of various transporters, overcoming limitations of conventional electrophysiology.
Area of Science:
- Membrane protein biophysics
- Electrophysiology
- Biochemical assay development
Background:
- Conventional electrophysiology faces challenges in characterizing low turnover or bacterial/intracellular transport proteins.
- Characterizing membrane transport proteins is crucial for understanding cellular functions and developing therapeutics.
Purpose of the Study:
- To present solid-supported membrane (SSM)-based electrophysiology as a sensitive, cell-free assay for electrogenic membrane protein characterization.
- To highlight the ability of SSM-based electrophysiology to determine kinetic properties of various transporters.
Main Methods:
- Purified proteins reconstituted into proteoliposomes or vesicles are adsorbed onto an SSM sensor.
- Rapid solution exchange applies substrates/ligands, and electrogenic activity generates a measurable current.
- High SSM stability allows cumulative measurements under varied conditions.
Main Results:
- The technique successfully characterized approximately 100 different transporters, including symporters, exchangers, uniporters, and various pumps.
- Kinetic parameters such as EC50, IC50, Km, KD, and rate constants were determined.
- Instruments range from laboratory setups to high-throughput systems like SURFE2R 96SE for simultaneous measurements.
Conclusions:
- SSM-based electrophysiology is a versatile and sensitive method for functional and kinetic characterization of diverse electrogenic membrane proteins.
- This cell-free approach expands the scope of electrophysiological studies to challenging transporter types and facilitates drug screening.
Keywords:
ATPaseBilayerCell freeElectrophysiologyMembrane transportPumpSSMSURFE(2)RSURFERSolid-supported membraneTransport assayTransporterMore Related Videos
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