Protein S-glutathionylation confers cellular resistance to ferroptosis induced by glutathione depletion
Yi Ju,
Yuting Zhang,
Xiaolin Tian,
Nanbin Zhu,
Yufan Zheng,
Yiming Qiao,
Tao Yang,
Baolin Niu,
Xiaoyun Li,
Liu Yu,
Zhuolin Liu,
Yixuan Wu,
Yang Zhi,
Yinuo Dong,
Qingling Xu,
Xiaoming Yang,
Xuening Wang,
Xiaokai Wang,
Haiteng Deng,
Yimin Mao,
Xiaobo Li
Affiliations
Yi Ju
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Yuting Zhang
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Xiaolin Tian
MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China
Nanbin Zhu
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Yufan Zheng
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Yiming Qiao
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Tao Yang
Third Hospital of Shanxi Medical University, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Taiyuan, China
Baolin Niu
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Xiaoyun Li
Division of Gastroenterology and Hepatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, NHC Key Laboratory of Digestive Diseases, Shanghai Research Center of Fatty Liver Disease, Shanghai, China
Liu Yu
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Zhuolin Liu
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Yixuan Wu
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China
Yang Zhi
Division of Gastroenterology and Hepatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, NHC Key Laboratory of Digestive Diseases, Shanghai Research Center of Fatty Liver Disease, Shanghai, China
Yinuo Dong
Division of Gastroenterology and Hepatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, NHC Key Laboratory of Digestive Diseases, Shanghai Research Center of Fatty Liver Disease, Shanghai, China
Qingling Xu
Department of Hepatology, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, Fujian, China
Xiaoming Yang
NHC Key Laboratory of Metabolic Cardiovascular Diseases Research, Ningxia Medical University, Yinchuan, Ningxia, China
Xuening Wang
Third Hospital of Shanxi Medical University, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Taiyuan, China
Xiaokai Wang
Department of Vascular Surgery, Xuzhou First People's Hospital, Xuzhou, Jiangsu, China
Haiteng Deng
MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China
Yimin Mao
Division of Gastroenterology and Hepatology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, NHC Key Laboratory of Digestive Diseases, Shanghai Research Center of Fatty Liver Disease, Shanghai, China; Corresponding author.
Xiaobo Li
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Fudan University, Shanghai, China; Corresponding author.
Ferroptosis is one of the most critical biological consequences of glutathione depletion. Excessive oxidative stress, indicated by an elevated oxidized glutathione (GSSG)/reduced glutathione (GSH) ratio, is recognized as a key driver of ferroptosis. However, in glutathione depletion-induced ferroptosis, a marked decrease in total glutathione levels (including both GSH and GSSG) is frequently observed, yet its significance remains understudied. Protein S-glutathionylation (protein-SSG) levels are closely linked to the redox state and cellular glutathione pools including GSH and GSSG. To date, the role of protein-SSG during cell ferroptosis induced by glutathione depletion remains poorly understood. Here, we demonstrated that upregulation of CHAC1, a glutathione-degrading enzyme, acted as a key regulator of protein-SSG formation and exacerbated glutathione depletion-induced ferroptosis. This effect was observed in both in vitro and in vivo models, including erastin-induced ferroptosis across multiple cell lines and acetaminophen overdose-triggered ferroptosis in hepatocytes. Deficiency of CHAC1 resulted in increased glutathione pools, enhanced protein-SSG, improved liver function, and attenuation of hepatocyte ferroptosis upon acetaminophen challenge. These protective effects were reversed by CHAC1 overexpression. Using quantitative redox proteomics, we identified glutathione pool-sensitive S-glutathionylated proteins. As an important example, we discovered that ADP-ribosylation factor 6 (ARF6) was regulated by S-glutathionylation during glutathione depletion-induced ferroptosis. Our findings revealed that CHAC1 upregulation reduced the S-glutathionylation of ARF6, resulting in decreased ARF6 levels in lysosomes. This, in turn, enhanced the localization of the transferrin receptor (TFRC) on the cell membrane and increased transferrin uptake, ultimately compromising the protective role of ARF6 in ferroptosis induced by glutathione depletion. Targeting TFRC using GalNAc-siTfrc mitigated acetaminophen-induced liver injury in vivo. In conclusion, our study provide evidence that availability of glutathione pools affects protein S-glutathionylation and regulates protein functions to influence the process of ferroptosis, which opens an avenue to understanding the cell ferroptosis induced by glutathione depletion.
