Dystonia-associated forms of torsinA are deficient in ATPase activity

J Mol Neurosci. 2005;25(1):105-17. doi: 10.1385/JMN:25:1:105.

Abstract

Early-onset dystonia is caused by mutations in the torsinA protein, a putative member of the AAA+ class of ATPases. In this study we have evaluated the ATPase activity of bacterially expressed wild-type torsinA and its disease-associated mutant forms. Upon overexpression in Escherichia coli, recombinant torsinA proteins were accumulated as insoluble inclusion bodies and required refolding to become soluble and catalytically active. The refolded wild-type and mutant torsinA proteins were capable of hydrolyzing ATP, but their specific ATPase activities differed significantly. Deletions of the amino acid residues E302/303 and F323-Y328 resulted in a decrease of ATPase activity to approximately 35% and approximately 75% of the wild-type level, respectively. ATPase activity of wild-type and mutant torsinA proteins was influenced by factors that varied with cell stress, such as temperature, pH, and ionic strength, and was inhibited by sodium vanadate. Our results provide the first direct evidence for a role of torsinA as an active ATPase and suggest that the mutations in torsinA might affect normal functions of the protein by reducing its enzymatic activity.

MeSH terms

  • Adenosine Triphosphatases / antagonists & inhibitors
  • Adenosine Triphosphatases / metabolism*
  • Adenosine Triphosphate / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Dystonia Musculorum Deformans / genetics*
  • Enzyme Stability
  • Humans
  • Molecular Chaperones / chemistry
  • Molecular Chaperones / genetics*
  • Molecular Chaperones / metabolism*
  • Molecular Sequence Data
  • Mutation
  • Protein Conformation
  • Protein Folding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Molecular Chaperones
  • Recombinant Proteins
  • TOR1A protein, human
  • Adenosine Triphosphate
  • Adenosine Triphosphatases