The P20R mutation of αB-crystallin diminishes its anti-apoptotic activity in human lens epithelial cells

Biochem Biophys Res Commun. 2017 Jan 29;483(1):463-467. doi: 10.1016/j.bbrc.2016.12.121. Epub 2016 Dec 19.

Abstract

αB-crystallin acts as an anti-apoptosis protein in human lens epithelial (HLE) cells. We recently identified a missense mutation in αB-crystallin that changes proline 20 to an arginine (P20R) in a Chinese family with autosomal dominant congenital posterior polar cataract. The impact of the P20R mutation on the anti-apoptosis function remains unclear. To explore the anti-apoptotic activity of αB-crystallin wild type (αB-wt) and its P20R mutant under oxidative stress, HLE cells were transfected with αB-wt and αB-P20R constructs and expression was measured by western blotting. Flow cytometry and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP digoxigenin nick end-labelling (TUNEL) staining were performed to investigate apoptosis. We found that αB-wt performed a dominant role in inhibiting stress-induced apoptosis, but this function was impeded in cells expressing αB-P20R. The P20R mutant of αB-crystallin exhibits diminished anti-apoptotic activity compared with the native protein.

Keywords: Anti-apoptosis; Point mutation; αB-crystallin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Apoptosis / genetics*
  • Arginine / genetics
  • Cataract / genetics
  • Cataract / pathology
  • Cells, Cultured
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Flow Cytometry
  • Humans
  • In Situ Nick-End Labeling
  • Lens, Crystalline / cytology*
  • Mutation, Missense
  • Proline / genetics
  • alpha-Crystallin B Chain / genetics*
  • alpha-Crystallin B Chain / metabolism

Substances

  • alpha-Crystallin B Chain
  • Arginine
  • Proline