In-cell NMR in E. coli to monitor maturation steps of hSOD1

PLoS One. 2011;6(8):e23561. doi: 10.1371/journal.pone.0023561. Epub 2011 Aug 24.

Abstract

In-cell NMR allows characterizing the folding state of a protein as well as posttranslational events at molecular level, in the cellular context. Here, the initial maturation steps of human copper, zinc superoxide dismutase 1 are characterized in the E. coli cytoplasm by in-cell NMR: from the apo protein, which is partially unfolded, to the zinc binding which causes its final quaternary structure. The protein selectively binds only one zinc ion, whereas in vitro also the copper site binds a non-physiological zinc ion. However, no intramolecular disulfide bridge formation occurs, nor copper uptake, suggesting the need of a specific chaperone for those purposes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoproteins / metabolism
  • Cell Extracts
  • Cysteine / metabolism
  • Cytoplasm / metabolism
  • Escherichia coli / metabolism*
  • Humans
  • Magnetic Resonance Spectroscopy
  • Oxidation-Reduction
  • Protein Binding
  • Protein Denaturation
  • Protein Folding
  • Protein Processing, Post-Translational*
  • Protein Structure, Quaternary
  • Superoxide Dismutase / chemistry
  • Superoxide Dismutase / metabolism*
  • Superoxide Dismutase-1
  • Tryptophan / metabolism
  • Zinc / metabolism

Substances

  • Apoproteins
  • Cell Extracts
  • SOD1 protein, human
  • Tryptophan
  • Superoxide Dismutase
  • Superoxide Dismutase-1
  • Zinc
  • Cysteine