Refolding studies using pressure: the folding landscape of lysozyme in the pressure-temperature plane

Biochim Biophys Acta. 2006 Mar;1764(3):497-505. doi: 10.1016/j.bbapap.2006.01.016. Epub 2006 Feb 13.

Abstract

Refolding of hen egg white lysozyme after pressure unfolding was measured by FTIR spectroscopy. The high-pressure treatment was found to be useful for unfolding/refolding studies because pressure acts against aggregation, and therefore no irreversible aggregation takes place during the pressure treatment. After the release of the pressure, folding intermediate structures were found which were formed during the decompression of the lysozyme. These were aggregation prone when heated, as indicated by their lower stability against aggregation. The intermediates were only formed if the protein was unfolded, subdenaturing pressures could not populate these intermediates. We introduced the notion of a superfunnel to describe the free energy landscape of interacting polypeptide chains. This can explain the propensity of folding intermediates to aggregate. A possible Gibbs-free energy landscape for lysozyme was constructed for the whole pressure-temperature plane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Crystallography, X-Ray
  • Muramidase / chemistry*
  • Pressure
  • Protein Binding*
  • Protein Conformation
  • Spectroscopy, Fourier Transform Infrared
  • Temperature

Substances

  • hen egg lysozyme
  • Muramidase