Glycine-alanine repeats impair proper substrate unfolding by the proteasome

Martin A Hoyt, Judith Zich, Junko Takeuchi, Mingsheng Zhang, Cedric Govaerts, Philip Coffino

Research output: Contribution to journalArticlepeer-review


Proteasome ATPases unravel folded proteins. Introducing a sequence containing only glycine and alanine residues (GAr) into substrates can impair their digestion. We previously proposed that a GAr interferes with the unfolding capacity of the proteasome, leading to partial degradation of products. Here we tested that idea in several ways. Stabilizing or destabilizing a folded domain within substrate proteins changed GAr-mediated intermediate production in the way predicted by the model. A downstream folded domain determined the sites of terminal proteolysis. The spacing between a GAr and a folded domain was critical for intermediate production. Intermediates containing a GAr did not remain associated with proteasomes, excluding models whereby retained GAr-containing proteins halt further processing. The following model is supported: a GAr positioned within the ATPase ring reduces the efficiency of coupling between nucleotide hydrolysis and work performed on the substrate. If this impairment takes place when unfolding must be initiated, insertion pauses and proteolysis is limited to the portion of the substrate that has already entered the catalytic chamber of the proteasome.
Original languageEnglish
Pages (from-to)1720-9
Number of pages10
JournalEMBO Journal
Issue number8
Publication statusPublished - 19 Apr 2006


  • Alanine
  • Animals
  • Glycine
  • Hydrolysis
  • Mice
  • Models, Molecular
  • Mutation
  • Ornithine Decarboxylase
  • Proteasome Endopeptidase Complex
  • Protein Folding
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae
  • Substrate Specificity
  • Tetrahydrofolate Dehydrogenase


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