Cobalt- and nickel-binding property of cullin-2

Biochem Biophys Res Commun. 2002 Jan 11;290(1):294-9. doi: 10.1006/bbrc.2001.6207.

Abstract

Treatment with divalent metal ions such as cobalt (Co(2+)) or nickel (Ni(2+)) result in the stabilization of hypoxia-inducible factor-1alpha (HIF1alpha). Recently, HIF1alpha was shown to be ubiquitinated by an E3-ligase complex and be subsequently targeted for proteasomal degradation. In this study, we demonstrated that Co(2+) and Ni(2+) specifically bind to cullin-2. Mutant analysis revealed that cullin-2 possesses at least three sites for the binding. Furthermore, fluorescence spectroscopy revealed that only Co(2+) and Ni(2+) have the binding activity to cullin-2, but other metal ions, including Cu(2+), Ca(2+), Mg(2+), Mn(2+), and Zn(2+), did not. Finally, we found that Co(2+) and Ni(2+) do not bind to any components of the E3-ligase other than cullin-2, suggesting that cullin-2 is a key target of Co(2+) and Ni(2+). Interestingly, Co(2+) did not affect the complex formation of the ligase, suggesting that the metal binding to cullin-2 affects the function, but not the assembly of the E3-ligase.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics
  • Animals
  • Binding Sites
  • Blotting, Western
  • COS Cells
  • Cell Cycle Proteins / chemistry*
  • Cell Cycle Proteins / metabolism*
  • Cell Line
  • Cobalt / chemistry*
  • Cobalt / metabolism
  • Cullin Proteins*
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Endothelium, Vascular / cytology
  • Genetic Vectors
  • Glutathione Transferase / chemistry
  • Glutathione Transferase / metabolism
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Ligases / metabolism
  • Magnesium / chemistry
  • Manganese / chemistry
  • Mutation
  • Nickel / chemistry*
  • Nickel / metabolism
  • Plasmids
  • Precipitin Tests
  • Protein Binding
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Spectrometry, Fluorescence
  • Transcription Factors / chemistry
  • Transcription, Genetic
  • Transfection
  • Ubiquitin-Protein Ligases
  • Umbilical Veins / cytology
  • Zinc / chemistry

Substances

  • CUL2 protein, human
  • Cell Cycle Proteins
  • Cullin Proteins
  • DNA, Complementary
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Cobalt
  • Manganese
  • Nickel
  • Ubiquitin-Protein Ligases
  • Glutathione Transferase
  • Ligases
  • Magnesium
  • Zinc