The identification of a reaction site of glutathione mixed-disulphide formation on γS-crystallin in human lens

Abstract

The glutathionylation of human lens proteins was examined by Western-blot analysis with an anti-GSH antibody and scanning. Several different glutathionylated proteins were observed, and a 47 kDa band was of particular interest. This band did not appear after SDS/PAGE under reducing conditions, suggesting that it was a glutathionylated fraction. The 47 kDa band was found principally in the outer part of the lens, the cortex, but not in the lens nucleus where older proteins are present. The 47 kDa component was composed of βB1-, βB2- and γS-crystallin, with the γS-crystallin having glutathione bound at Cys-82 and at Cys-22, Cys-24 or Cys-26. We conclude that when glutathione becomes bound to γS-crystallin, it causes it to bind in turn to the β-crystallin polypeptides to form a dimer.