Identification of an artificial peptide motif that binds and stabilizes reduced human DJ-1

J Struct Biol. 2011 Dec;176(3):414-8. doi: 10.1016/j.jsb.2011.08.011. Epub 2011 Aug 27.


Although the precise biochemical function of DJ-1 remains unclear, it has been found to exert cytoprotective activity against oxidative stress. Cys106 is central to this function since it has a distinctly low pK(a) rendering it extremely susceptible for oxidation. This characteristic, however, also poses a severe hindrance to obtain reduced DJ-1 for in vitro investigation. We have developed an approach to produce recombinant human DJ-1 in its reduced form as a bona fide basis for exploring the redox capacities of the protein. We solved the crystal structure of this DJ-1 at 1.56Å resolution, allowing us to capture Cys106 in the reduced state for the first time. The dimeric structure reveals one molecule of DJ-1 in its reduced state while the other exhibits the characteristics of a mono-oxygenated cysteine. Comparison with previous structures indicates the absence of redox dependent global conformational changes in DJ-1. The capture of reduced Cys106 is facilitated by stabilization within the putative active site achieved through a glutamate side chain. This side chain is provided by a crystallographic neighbor as part of a 'Leu-Glu' motif, which was added to the C-terminus of DJ-1. In the structure this motif binds DJ-1 in close proximity to Cys106 through extended hydrophilic and hydrophobic interactions depicting a distinct binding pocket, which can serve as a basis for compound development targeting DJ-1.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Motifs
  • Binding Sites
  • Catalytic Domain
  • Crystallography, X-Ray
  • Cysteine / chemistry*
  • Glutamine / chemistry
  • Humans
  • Intracellular Signaling Peptides and Proteins / chemistry*
  • Leucine / chemistry
  • Oncogene Proteins / chemistry*
  • Oxidation-Reduction
  • Oxidative Stress
  • Peptides / chemical synthesis
  • Peptides / chemistry*
  • Protein Binding
  • Protein Deglycase DJ-1
  • Protein Stability
  • Reactive Oxygen Species / chemistry
  • Recombinant Proteins / chemistry


  • Intracellular Signaling Peptides and Proteins
  • Oncogene Proteins
  • Peptides
  • Reactive Oxygen Species
  • Recombinant Proteins
  • Glutamine
  • PARK7 protein, human
  • Protein Deglycase DJ-1
  • Leucine
  • Cysteine