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Ion Conductivity of SecYEG Engaged in Translocation

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SecYEG gating is voltage-sensitive.

Linz Winter Workshop 2015

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International Workshop: Permeation & Gating of Ion Channels, June 2014 in Austria

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Christian Doppler Labor: Nanoskopische Methoden in der Biophysik

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Membrane Transport, Peter Pohl

The research foci include transport of water, protons, other small molecules, and proteins through biological membranes. We thus exploit (i) primary cells or tissues, (ii) cultured cells that are genetically modified to overexpress the protein of interest or (iii) planar lipid bilayers or lipid vesicles both reconstituted with the purified membrane transporter. Purification occurs by overexpressing E.coli cells or yeast cells using affinity and size-exclusion chromatography.
      Our work aims to unravel molecular transport mechanisms. We specifically want to understand (i) the major determinants of single file water transport in potassium channels, aquaporins, artificial nanopores, (ii) the principles of water flux through secondary active transporters, (iii) the structural-functional relationship which allows the protein translocase to facilitate the passage of large molecules while maintaining the membrane barrier for small molecules, (iv) the mechanisms of lateral proton migration between two membrane proteins.

The proteins under investigation are genetically or chemically modified to test the functional importance of certain residues and to introduce fluorescent labels. We monitor protein function by implementing a wide variety of methods including: recordings of current through single channels and through channel ensembles, streaming and boundary potential measurements, particle electrophoresis, stopped- flow spectroscopy, fluorescence correlation spectroscopy, fluorimetry, confocal fluorescence microscopy, and scanning electrochemical microscopy.

 
 

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