Cataract-associated P23T γD-crystallin retains a native-like fold in amorphous-looking aggregates formed at physiological pH

Nat Commun. 2017 May 5:8:15137. doi: 10.1038/ncomms15137.

Abstract

Cataracts cause vision loss through the large-scale aggregation of eye lens proteins as a result of ageing or congenital mutations. The development of new treatments is hindered by uncertainty about the nature of the aggregates and their mechanism of formation. We describe the structure and morphology of aggregates formed by the P23T human γD-crystallin mutant associated with congenital cataracts. At physiological pH, the protein forms aggregates that look amorphous and disordered by electron microscopy, reminiscent of the reported formation of amorphous deposits by other crystallin mutants. Surprisingly, solid-state NMR reveals that these amorphous deposits have a high degree of structural homogeneity at the atomic level and that the aggregated protein retains a native-like conformation, with no evidence for large-scale misfolding. Non-physiological destabilizing conditions used in many in vitro aggregation studies are shown to yield qualitatively different, highly misfolded amyloid-like fibrils.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cataract / congenital
  • Cataract / genetics*
  • Humans
  • Hydrogen-Ion Concentration
  • In Vitro Techniques
  • Magnetic Resonance Spectroscopy
  • Microscopy, Electron, Transmission
  • Mutation
  • Protein Aggregates*
  • Protein Aggregation, Pathological / genetics*
  • Protein Aggregation, Pathological / metabolism
  • Protein Aggregation, Pathological / pathology
  • Protein Conformation
  • Protein Folding
  • gamma-Crystallins / genetics*
  • gamma-Crystallins / metabolism
  • gamma-Crystallins / ultrastructure

Substances

  • CRYGD protein, human
  • Protein Aggregates
  • gamma-Crystallins