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X-Ray Crystallography of Carbon Monoxide Dehydrogenases

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Metalloproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1876))

Abstract

Carbon monoxide dehydrogenases (CODHs) are central players in the biogeochemical carbon monoxide (CO) cycle and have been extensively studied from the ecological level to the structural/molecular level. Of the two types of CODHs, the oxygen-tolerant CODHs use a bimetallic [CuSMo(=O)OH] center connected to the protein via a pyranopterin cofactor, whereas the oxygen-sensitive CODHs contain a [NiFe4S4-OHx]-cluster. Despite the fact that we have a basic understanding of how both types of CODHs use distinct active sites to catalyze the oxidation of CO with water to CO2, two protons, and two electrons (a reversible reaction in the cases of the oxygen-sensitive CODHs), many questions remain unanswered, especially concerning the electronic structures of the intermediate states. Since these states will likely be only revealed by the interplay of experimental and theoretical methods, there is a need to obtain accurate descriptions of the active site architectures in various states and, consequently, a need to generate crystals with good diffraction quality and collect data at element-specific wavelengths in order to determine the identity of elements in the case of mixed states. This chapter provides a description of the general working protocols for the crystallization and structural analysis of Cu,Mo-CODH and Ni,Fe-CODH that facilitates the mechanistic investigations of these important metalloenzymes.

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Acknowledgments

The authors are supported by the German funding agency DFG project grants (DO-785/1, DO-787/5, DO-785/6) and the Cluster of Excellence “Unifying Concepts in Catalysis – UniCat” (EXC 314).

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Authors and Affiliations

  1. Institute of Biology, Structural Biology and Biochemistry, Humboldt-Universität zu Berlin, Berlin, Germany

    Jae-Hun Jeoung, Berta M. Martins & Holger Dobbek

Authors
  1. Jae-Hun Jeoung
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  2. Berta M. Martins
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  3. Holger Dobbek
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Corresponding author

Correspondence to Holger Dobbek .

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Editors and Affiliations

  1. Department of Molecular Biology and Biochemistry, University of California-Irvine, Irvine, CA, USA

    Yilin Hu

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Jeoung, JH., Martins, B.M., Dobbek, H. (2019). X-Ray Crystallography of Carbon Monoxide Dehydrogenases. In: Hu, Y. (eds) Metalloproteins. Methods in Molecular Biology, vol 1876. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8864-8_11

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  • DOI: https://doi.org/10.1007/978-1-4939-8864-8_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8863-1

  • Online ISBN: 978-1-4939-8864-8

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