A biophysical analysis of the tetratricopeptide repeat-rich mitochondrial import receptor, Tom70, reveals an elongated monomer that is inherently flexible, unstable, and unfolds via a multistate pathway

J Biol Chem. 2004 Nov 5;279(45):46448-54. doi: 10.1074/jbc.M405639200. Epub 2004 Aug 16.

Abstract

Proteins destined for all submitochondrial compartments are translocated across the outer mitochondrial membrane by the TOM (translocase of the outer membrane) complex, which consists of a number of specialized receptor subunits that bind mitochondrial precursor proteins for delivery into the translocation channel. One receptor, Tom70, binds large, hydrophobic mitochondrial precursors. The current model of Tom70-mediated import involves multiple dimers of the receptor recognizing a single molecule of substrate. Here we show via a battery of biophysical and spectroscopic techniques that the cytosolic domain of Tom70 is an elongated monomer. Thermal and urea-induced denaturation revealed that the receptor, which unfolds via a multistate pathway, is a relatively unstable molecule undergoing major conformational change at physiological temperatures. The data suggest that the malleability of the monomeric Tom70 receptor is an important factor in mitochondrial import.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Binding Sites
  • Circular Dichroism
  • Cytosol / metabolism
  • DNA / metabolism
  • Dimerization
  • Dose-Response Relationship, Drug
  • Electrophoresis, Polyacrylamide Gel
  • Hot Temperature
  • Ligands
  • Membrane Proteins / physiology*
  • Mitochondria / metabolism*
  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Precursor Protein Import Complex Proteins
  • Protein Conformation
  • Protein Denaturation
  • Protein Folding
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / physiology*
  • Spectrometry, Fluorescence
  • Temperature
  • Thermodynamics
  • Ultracentrifugation
  • Urea / pharmacology

Substances

  • Ligands
  • Membrane Proteins
  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Precursor Protein Import Complex Proteins
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • TOM70 protein, S cerevisiae
  • Urea
  • DNA