Pressure-jump NMR study of dissociation and association of amyloid protofibrils

J Mol Biol. 2005 Jun 24;349(5):916-21. doi: 10.1016/j.jmb.2005.04.010.

Abstract

The dissociation and reassociation processes of amyloid protofibrils initiated by pressure-jump have been monitored with real-time (1)H NMR spectroscopy using an intrinsically denatured disulfide-deficient variant of hen lysozyme. Upon pressure-jump up to 2 kbar, the matured protofibrils grown over several months become fully dissociated into monomers within a few days. Upon pressure-jump down to 30 bar, the dissociated monomers immediately start reassociating. The association and dissociation cycle can be repeated reproducibly by alternating pressure, establishing a notion that the protofibril formation is simply a slow kinetic process toward thermodynamic equilibrium. The outstanding simplicity and effectiveness of pressure in controlling the protofibril formation opens a new route for investigating mechanisms of amyloid fibril-forming reactions. The noted variation in the pressure-induced dissociation rate with the progress of the association reaction suggests multiple mechanisms for the elongation of the protofibril. The disulfide-deficient hen lysozyme offers a particularly simple model system for thermodynamic and kinetic studies of protofibril formation as well as for screening drugs for amyloidosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid / physiology*
  • Amyloid / ultrastructure
  • Animals
  • Chickens
  • Microscopy, Atomic Force
  • Muramidase / physiology*
  • Muramidase / ultrastructure
  • Nuclear Magnetic Resonance, Biomolecular
  • Pressure
  • Protein Denaturation
  • Thermodynamics

Substances

  • Amyloid
  • Muramidase