In vitro characterization of PGAM5, Bcl-XL, and Bax: mitochondrial proteins regulating mitochondrion dynamic and cell death
Zhi Zhang1, Kaili Ma3, Jianyu Feng1,3, Yushan Zhu3, Quan Chen3,4, and Jialing Lin1,2 1Department of Biochemistry and Molecular Biology, 2Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, USA; 3State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, Tianjin, China; 4State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
PGAM5 is a mitochondrial serine/threonine phosphatase that interacts with proteins of the Bcl-2 family including anti-apoptotic Bcl-XL and pro-apoptotic Bax to regulate mitofission, mitophagy, and apoptosis. How these proteins interact with each other and how their interactions regulate the structure and function of each protein are unclear. Here, we synthesize these proteins in vitro to study their localization to and interactions at mitochondria using fractionation, protease digestion, site-specific crosslink, and other biochemical methods. We assess the impact of their interactions on the conformation, self-association, and function of each protein using fluorescence, small angle X-ray scattering, and other biophysical methods. We will report our findings and their implications to the molecular mechanisms by which these proteins regulate mitochondrial dynamics and cell death. (This work is supported by NIH grants P20GM103640 and R01GM062964 to J.L., and National Natural Science Foundation of China grant 31520103904 to Q.C.)
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