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Anaerobic MicroScale Thermophoresis reveals the Redox dependency of ferredoxin in mitochondrial Fe/S biogenesis

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4 7 Stehling, O. & Lill, R. The role of mitochondria in cellular iron-sulfur protein biogenesis: mechanisms, connected processes, and diseases. Cold Spring Harbor perspectives in biology 5, a011312, doi:10.1101/cshperspect.a011312 (2013). 8 Beilschmidt, L. K. & Puccio, H. M. Mammalian Fe-S cluster biogenesis and its implication in disease. Biochimie, doi:10.1016/j.biochi.2014.01.009 (2014). 9 Tsai, C. L. & Barondeau, D. P. Human frataxin is an allosteric switch that activates the Fe-S cluster biosynthetic complex. Biochemistry 49, 9132-9139, doi:10.1021/bi1013062 (2010). 10 Shi, Y., Ghosh, M., Kovtunovych, G., Crooks, D. R. & Rouault, T. A. Both human ferredoxins 1 and 2 and ferredoxin reductase are important for iron- sulfur cluster biogenesis. Biochimica et biophysica acta 1823, 484-492, doi:10.1016/j.bbamcr.2011.11.002 (2012). 11 Zheng, L., Cash, V. L., Flint, D. H. & Dean, D. R. Assembly of iron-sulfur clusters. Identification of an iscSUA-hscBA-fdx gene cluster from Azotobacter vinelandii. J. Biol. Chem. 273, 13264-13272. (1998). 12 Tokumoto, U. & Takahashi, Y. Genetic Analysis of the isc Operon in Escherichia coli Involved in the Biogenesis of Cellular Iron-Sulfur Proteins. J. Biochem. (Tokyo) 130, 63-71. (2001) 13 Kim, J. H., Frederick, R. O., Reinen, N. M., Troupis, A. T. & Markley, J. L. [2Fe-2S]-ferredoxin binds directly to cysteine desulfurase and supplies an electron for iron-sulfur cluster assembly but is displaced by the scaffold protein or bacterial frataxin. J Am Chem Soc 135, 8117-8120, doi:10.1021/ja401950a (2013). 14 Mühlenhoff, U., Richter, N., Pines, O., Pierik, A. J. & Lill, R. Specialized function of yeast Isa1 and Isa2 proteins in the maturation of mitochondrial [4Fe-4S] proteins. J Biol Chem 286, 41205-41216, doi:10.1074/jbc.M111.296152 (2011). 15 Wienken, C. J., Baaske, P., Rothbauer, U., Braun, D. & Duhr, S. Protein-binding assays in biological liquids using microscale thermophoresis. Nat Commun 1, 100, doi:10.1038/ncomms1093 (2010). 16 Gerber, J., Neumann, K., Prohl, C., Mühlenhoff, U. & Lill, R. The yeast scaffold proteins Isu1p and Isu2p are required inside mitochondria for maturation of cytosolic Fe/S proteins. Mol. Cell Biol. 24, 4848-4857. (2004). @ 2015 NanoTemper Technologies GmbH

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