Heterologous Production and Characterization of Multiheme Cytochrome c Enzymes
Final Report Abstract
Multiheme Cytochromes c are a diverse family of electron carriers and redox enzymes that play a central role in several metabolic pathways within the biogeochemical nitrogen cycles of nitrogen, sulfur and iron. Their heme cofactors pack into conserved interaction motifs that allow for strong electronic coupling and efficient electron transfer. In this project, various multiheme cytochromes c have been recombinantly produced in either Escherichia coli or Wolinella succinogenes cells using sophisticated gene expression and cytochrome c biogenesis systems. The studied cytochromes c included diheme cytochrome c peroxidases, pentaheme cytochrome c oxidoreductases of the NrfA family, octaheme cytochrome c oxidoreductases of the Hao and MccA families and the nonaheme cytochrome c OcwA involved in metal reduction by Gram-positives. The cytochromes were purified and characterized in terms of enzyme activities, substrate/product range, spectroscopic features, high-resolution crystal structures and evolutionary implications. The results contributed to our understanding of the structure/function relationships of periplasmic/extracellular cytochrome c-dependent electron transport networks that are crucial for many environmentally significant processes.
Publications
- (2011) Physiological function and catalytic versatility of bacterial multihaem cytochromes c involved in nitrogen and sulfur cycling. Biochem. Soc. Trans. 39, 1864-1870
Simon, J., Kern, M., Hermann, B., Einsle, O. & Butt, J.N.
(See online at https://doi.org/10.1042/BST20110713) - (2011) The oxidative and nitrosative stress defence network of Wolinella succinogenes: cytochrome c nitrite reductase mediates the stress response to nitrite, nitric oxide, hydroxylamine and hydrogen peroxide. Environ. Microbiol. 13, 2478-2494
Kern, M., Volz, J. & Simon, J.
(See online at https://doi.org/10.1111/j.1462-2920.2011.02520.x) - (2012) Enzyme or Electrode? Structure 20, 1132-1134
Einsle, O.
(See online at https://doi.org/10.1016/j.str.2012.06.005) - (2012) MacA is a second cytochrome c Peroxidase in Geobacter sulfurreducens. Biochemistry 51, 2747-2456
Seidel, J., Hoffmann, M., Ellis, K.E., Seidel, A., Spatzal, T., Gerhardt, S., Elliott, S.J. & Einsle, O.
(See online at https://doi.org/10.1021/bi300249u) - (2013) Structure of the processive rubber oxygenase RoxA from Xanthomonas sp. Proc. Natl. Acad. Sci. USA 110, 13833-13838
Seidel, J., Schmitt, G., Hoffmann, M., Jendrossek, D. & Einsle, O.
(See online at https://doi.org/10.1073/pnas.1305560110) - (2014) The production of ammonia by multiheme cytochromes c. In: The Metal-Driven Biogeochemistry of Gaseous Compounds in the Environment, Eds P.M.H. Kroneck, M.E. Sosa Torres. Met. Ions Life Sci. 14, 211-236
Simon, J. & Kroneck, P.M.H.
(See online at https://doi.org/10.1007/978-94-017-9269-1_9) - (2015) Multiheme Peroxidases. In: Heme Peroxidases (Raven, E.L. & Dunford, H.B., eds.) RSC Publishing, Oxford, 113-132
Brausemann, A., Seidel, J., Wüst, A. & Einsle, O.
(See online at https://doi.org/10.1039/9781782622628-00113) - (2015) Resolution of key roles for the distal pocket histidine in cytochrome c nitrite reductases. J. Am. Chem. Soc. 137, 3059-3068
Lockwood, C.W.J., Burlat, B., Cheesman, M.R., Kern, M., Simon, J., Clarke, T.A., Richardson, D.J. & Butt, J.N.
(See online at https://doi.org/10.1021/ja512941j) - (2015) The octahaem MccA is a haem c–copper sulfite reductase. Nature 520, 706-709
Hermann, B., Kern, M., La Pietra, L., Simon, J. & Einsle, O.
(See online at https://doi.org/10.1038/nature14109) - (2017) Epsilonproteobacterial hydroxylamine oxidoreductase (εHao): Characterization of a 'missing link' in the multihaem cytochrome c family. Mol. Microbiol. 105, 127-138
Haase, D., Hermann, B., Einsle, O. & Simon, J.
(See online at https://doi.org/10.1111/mmi.13690) - (2017) NMR studies of the interaction between inner membrane-associated and periplasmic cytochromes from Geobacter sulfurreducens. FEBS Lett. 591, 1657-1666
Dantas, J.M., Brausemann, A., Einsle, O. & Salgueiro, C.A.
(See online at https://doi.org/10.1002/1873-3468.12695) - (2017) Structural and functional analysis of latex clearing protein (Lcp) provides insight into the enzymatic cleavage of rubber. Sci. Rep. 7, 6179
Ilcu, L., Röther, W., Birke, J., Brausemann, A., Einsle, O. & Jendrossek, D.
(See online at https://doi.org/10.1038/s41598-017-05268-2) - (2019) How Thermophilic Gram-Positive Organisms Perform Extracellular Electron Transfer: Characterization of the Cell Surface Terminal Reductase OcwA. mBio 10, e01210-e01219
Costa, N.L., Hermann, B., Fourmond, V., Faustino, M.M., Teixeira, M., Einsle, O., Paquete, C.M. & Louro, R.O.
(See online at https://doi.org/10.1128/mBio.01210-19)
