Human γS-Crystallin-Copper Binding Helps Buffer against Aggregation Caused by Oxidative Damage

Biochemistry. 2020 Jun 30;59(25):2371-2385. doi: 10.1021/acs.biochem.0c00293. Epub 2020 Jun 12.

Abstract

Divalent metal cations can play a role in protein aggregation diseases, including cataract. Here we compare the aggregation of human γS-crystallin, a key structural protein of the eye lens, via mutagenesis, ultraviolet light damage, and the addition of metal ions. All three aggregation pathways result in globular, amorphous-looking structures that do not elongate into fibers. We also investigate the molecular mechanism underlying copper(II)-induced aggregation. This work was motivated by the observation that zinc(II)-induced aggregation of γS-crystallin is driven by intermolecular bridging of solvent-accessible cysteine residues, while in contrast, copper(II)-induced aggregation of this protein is exacerbated by the removal of solvent-accessible cysteines via mutation. Here we find that copper(II)-induced aggregation results from a complex mechanism involving multiple interactions with the protein. The initial protein-metal interactions result in the reduction of Cu(II) to Cu(I) with concomitant oxidation of γS-crystallin. In addition to the intermolecular disulfides that represent a starting point for aggregation, intramolecular disulfides also occur in the cysteine loop, a region of the N-terminal domain that was previously found to mediate the early stages of cataract formation. This previously unobserved ability of γS-crystallin to transfer disulfides intramolecularly suggests that it may serve as an oxidation sink for the lens after glutathione levels have become depleted during aging. γS-Crystallin thus serves as the last line of defense against oxidation in the eye lens, a result that underscores the chemical functionality of this protein, which is generally considered to play a purely structural role.

Publication types

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

MeSH terms

  • Copper / chemistry
  • Copper / metabolism*
  • Cysteine / chemistry
  • Disulfides / chemistry
  • Humans
  • Mutation
  • Oxidation-Reduction
  • Protein Binding
  • Protein Multimerization / drug effects*
  • Protein Multimerization / radiation effects
  • Ultraviolet Rays
  • gamma-Crystallins / chemistry
  • gamma-Crystallins / genetics
  • gamma-Crystallins / metabolism*

Substances

  • Disulfides
  • gamma-Crystallins
  • CRYGS protein, human
  • Copper
  • Cysteine