cytoplasmic and nuclear anti-apoptotic roles of αb-crystallin in retinal pigment epithelial cells细胞质和核αb-crystallin在视网膜色素上皮细胞的抗凋亡作用.pdf

Cytoplasmic and Nuclear Anti-Apoptotic Roles of aB- Crystallin in Retinal Pigment Epithelial Cells 1. 1. 2.¤ 2 2 Woo Jin Jeong , Jee Hyun Rho , Young Geol Yoon , Seung Hee Yoo , Na Young Jeong , Won 1 1 1 1 3 4 Yeol Ryu , Hee Bae Ahn , Woo Chan Park , Sae Heun Rho , Hee Seong Yoon , Yung Hyun Choi , Young Hyun Yoo2* 1 Department of Ophthalmology, Dong-A University College of Medicine, Seo-gu, Busan, Republic of Korea, 2 Department of Anatomy and Cell Biology, Dong-A University College of Medicine and Mitochondria Hub Regulation Center, Seo-gu, Busan, Republic of Korea, 3 Sungmo Eye Hospital, Inc., Haeundae-gu, Busan, Republic of Korea, 4 Department of Biochemistry and Research Institute of Oriental Medicine, Dongeui University College of Oriental Medicine, Busanjin-gu, Busan, Republic of Korea Abstract In addition to its well-characterized role in the lens, aB-crystallin performs other functions. Methylglyoxal (MGO) can alter the function of the basement membrane of retinal pigment epithelial (RPE) cells. Thus, if MGO is not efficiently detoxified, it can induce adverse reactions in RPE cells. In this study, we examined the mechanisms underlying the anti-apoptotic activity of aB-crystallin in the human retinal pigment epithelial cell line ARPE-19 following MGO treatment using various assays, including nuclear staining, flow cytometry, DNA electrophoresis, pulse field gel electrophoresis, western blot analysis, confocal microscopy and co-immunoprecipitation assays. To directly assess the role of phosphorylation of aB-crystallin, we used site-directed mutagenesis to convert