title: Functional refolding and characterization of two Tom40 isoforms from human mitochondria.
abstract: Tom40 proteins represent an essential class of molecules which facilitate translocation of unfolded proteins from the cytosol into the mitochondrial intermembrane space. They are part of a high-molecular mass complex that forms the protein-conducting channel in outer mitochondrial membranes. This study concerns the recombinant expression, purification and folding of amino-terminally truncated variants of the two human Tom40 isoforms for structural biology experiments. Both CD and FTIR secondary structure analysis revealed a dominant beta-sheet structure and a short alpha-helical part for both proteins together with a high thermal stability. Two secondary structure elements can be denatured independently. Reconstitution of the recombinant protein into planar lipid bilayers demonstrated ion channel activity similar to Tom40 purified from Neurospora crassa mitochondrial membranes, but conductivity fingerprints differ from the structurally closely related VDAC proteins.
authors: Biophysics Department, Institute of Biology, University of Stuttgart, Germany.
authors: Frauke
authors: F
authors: Mager
authors: Dennis
authors: D
authors: Gessmann
authors: Stephan
authors: S
authors: Nussberger
authors: Kornelius
authors: K
authors: Zeth
mesh_terms: D002942:Circular Dichroism; D004594:Electrophysiology; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D020033:Protein Isoforms / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D008051:Lipid Bilayers; D024101:Mitochondrial Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins