The effects of aB-crystallin on mitochondrial death pathway during hydrogen peroxide induced apoptosis

Abstract

aB-crystallin, a major small heat shock protein, has recently been shown to exert inhibitory effects on apoptosis, while the responsible mechanisms remain largely unknown. In the present study, we discovered that aB-crystallin protected mouse myoblast C2C12 cells against oxidative stress-induced apoptosis. During hydrogen peroxide-induced apoptosis, aBcrystallin showed that it decreased the redistribution level of phosphatidylserine (PS), reduced the release of cytochrome C and Smac/Diablo from mitochondria into cytoplasm, and decreased the cleavage of Bid. Interestingly, immunoprecipitation experiments with anti-aBcrystallin and anti-myc-tag antibodies demonstrated respectively an interaction between aBcrystallin and p53 during hydrogen peroxide induced apoptosis. Both the NH2-terminal and COOH-terminal regions of aB-crystallin could interact with p53, suggesting two domains of aB-crystallin are necessary for the interaction. Electrophoresis mobility shift assay (EMSA) and luciferase assay further demonstrated that aB-crystallin inhibited the upregulation of the DNA-binding, as well as the transactivation activity of p53 induced by hydrogen peroxide. Our results show that aB-crystallin has a protective role in oxidative stress induced apoptosis by interference with the mitochondrial death pathway.