Abstract
Native electrophoresis is a powerful tool to analyze the mitochondrial electron transport chain complexes (Cx) I–V and their assembly into supercomplexes. Valuable information regarding the composition and bioenergetic regulation in physiological and pathological conditions can be obtained. This chapter compares different types of native electrophoresis to analyze mitochondrial supercomplexes.
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References
Schagger H, von Jagow G (1991) Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 199(2):223–231
Wittig I, Karas M, Schagger H (2007) High resolution clear native electrophoresis for in-gel functional assays and fluorescence studies of membrane protein complexes. Mol Cell Proteomics 6(7):1215–1225. https://doi.org/10.1074/mcp.M700076-MCP200
Grandier-Vazeille X, Guerin M (1996) Separation by blue native and colorless native polyacrylamide gel electrophoresis of the oxidative phosphorylation complexes of yeast mitochondria solubilized by different detergents: specific staining of the different complexes. Anal Biochem 242(2):248–254. https://doi.org/10.1006/abio.1996.0460
Cuillerier A, Burelle Y (2019) Hybrid clear/blue native electrophoresis for the separation and analysis of mitochondrial respiratory chain supercomplexes. J Vis Exp (147). https://doi.org/10.3791/59294
Newell C, Khan A, Sinasac D et al (2019) Hybrid gel electrophoresis using skin fibroblasts to aid in diagnosing mitochondrial disease. Neurol Genet 5(3):e336. https://doi.org/10.1212/NXG.0000000000000336
Beutner G, Porter GA Jr (2017) Analyzing supercomplexes of the mitochondrial electron transport chain with native electrophoresis, in-gel assays, and electroelution. J Vis Exp (124). https://doi.org/10.3791/55738
Wittig I, Carrozzo R, Santorelli FM et al (2007) Functional assays in high-resolution clear native gels to quantify mitochondrial complexes in human biopsies and cell lines. Electrophoresis 28(21):3811–3820. https://doi.org/10.1002/elps.200700367
Wittig I, Schagger H (2009) Native electrophoretic techniques to identify protein-protein interactions. Proteomics 9(23):5214–5223. https://doi.org/10.1002/pmic.200900151
Wittig I, Beckhaus T, Wumaier Z et al (2010) Mass estimation of native proteins by blue native electrophoresis: principles and practical hints. Mol Cell Proteomics 9(10):2149–2161. https://doi.org/10.1074/mcp.M900526-MCP200
Schagger H (2001) Blue-native gels to isolate protein complexes from mitochondria. Methods Cell Biol 65:231–244
Beutner G, Alanzalon RE, Porter GA Jr (2017) Cyclophilin D regulates the dynamic assembly of mitochondrial ATP synthase into synthasomes. Sci Rep 7(1):14488. https://doi.org/10.1038/s41598-017-14795-x
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Beutner, G., Porter, G.A. (2021). Native Gel Electrophoresis and Immunoblotting to Analyze Electron Transport Chain Complexes. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2276. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1266-8_7
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DOI: https://doi.org/10.1007/978-1-0716-1266-8_7
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