The crystal structure of human ferroptosis suppressive protein 1 in complex with flavin adenine dinucleotide and nicotinamide adenine nucleotide
Shijian Feng,
Xiaofang Huang,
Dan Tang,
Xiaoyu Liu,
Liang Ouyang,
Dehua Yang,
Kunjie Wang,
Banghua Liao,
Shiqian Qi
Affiliations
Shijian Feng
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Xiaofang Huang
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Dan Tang
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Xiaoyu Liu
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Liang Ouyang
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Dehua Yang
The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences Shanghai China
Kunjie Wang
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Banghua Liao
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Shiqian Qi
Department of Urology and Institute of Urology (Laboratory of Reconstructive Urology) State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, College of Life Sciences, Sichuan University Chengdu China
Abstract Ferroptosis is a recently discovered form of regulated cell death characterized by its distinct dependence on iron and the peroxidation of lipids within cellular membranes. Ferroptosis plays a crucial role in physiological and pathological situations and has attracted the attention of numerous scientists. Ferroptosis suppressive protein 1 (FSP1) is one of the main regulators that negatively regulates ferroptosis through the GPX4‐independent FSP1–CoQ10–NAD(P)H axis and is a potential therapeutic target for ferroptosis‐related diseases. However, the crystal structure of FSP1 has not been resolved, which hinders the development of therapeutic strategies targeting FSP1. To unravel this puzzle, we purified the human FSP1 (hFSP1) protein using the baculovirus eukaryotic cell expression system and solved its crystal structure at a resolution of 1.75 Å. Furthermore, we evaluated the oxidoreductase activity of hFSP1 with NADH as the substrate and identified E156 as the key amino acid in maintaining hFSP1 activity. Interestingly, our results indicated that hFSP1 exists and functions in a monomeric state. Mutagenesis analysis revealed the critical role of the C‐terminal domain in the binding of substrate. These findings significantly enhance our understanding of the functional mechanism of FSP1 and provide a precise model for further drug development.
