In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants

Nat Commun. 2014 Nov 27:5:5502. doi: 10.1038/ncomms6502.

Abstract

Mutations in the superoxide dismutase 1 (SOD1) gene are related to familial cases of amyotrophic lateral sclerosis (fALS). Here we exploit in-cell NMR to characterize the protein folding and maturation of a series of fALS-linked SOD1 mutants in human cells and to obtain insight into their behaviour in the cellular context, at the molecular level. The effect of various mutations on SOD1 maturation are investigated by changing the availability of metal ions in the cells, and by coexpressing the copper chaperone for SOD1, hCCS. We observe for most of the mutants the occurrence of an unstructured SOD1 species, unable to bind zinc. This species may be a common precursor of potentially toxic oligomeric species, that are associated with fALS. Coexpression of hCCS in the presence of copper restores the correct maturation of the SOD1 mutants and prevents the formation of the unstructured species, confirming that hCCS also acts as a molecular chaperone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyotrophic Lateral Sclerosis / genetics*
  • Amyotrophic Lateral Sclerosis / metabolism
  • Copper / metabolism
  • HEK293 Cells
  • Humans
  • Molecular Chaperones / metabolism*
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Folding
  • Superoxide Dismutase / genetics*
  • Superoxide Dismutase / metabolism
  • Superoxide Dismutase-1
  • Zinc / metabolism

Substances

  • CCS protein, human
  • Molecular Chaperones
  • SOD1 protein, human
  • Copper
  • Superoxide Dismutase
  • Superoxide Dismutase-1
  • Zinc