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Magnetic Circular Dichroism Spectroscopy of Metalloproteins

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Metalloproteins

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

Abstract

Metals and metal clusters in proteins typically serve as important structural/functional motifs. Because of this reason, there is a wide range of techniques that specifically probe the structure and energy levels of metals in metalloproteins. One technique, magnetic circular dichroism (MCD) spectroscopy, is the focus of this chapter. MCD spectroscopy monitors the circular dichroism spectrum induced by a magnetic field and is an effective way of obtaining electronic and structural information of paramagnetic metal ions or clusters. The basic methodology of this technique is discussed along with examples of how MCD spectroscopy can be used to elucidate typical metal clusters in proteins. Special emphasis is placed on iron–sulfur (FeS) clusters.

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References

  1. Piepho SB, Schatz PN (1983) Group theory in spectroscopy with applications to magnetic circular dichroism. John Wiley & Sons, New York

    Google Scholar 

  2. Thompson AJ, Cheesman MR, George SJ (1993) Variable-temperature magentic circular dichroism. In: Riordan JF, Vallee BL (eds) Metallobiochemistry: part C, vol 226. Academic Press, San Diego, CA, pp 199–231

    Google Scholar 

  3. Johnson MK (2000) CD and MCD spectroscopy. In: Que LJ (ed) Physical methods in bioinorganic chemistry: spectroscopy and magnetism. University Science Books, Sausalito, pp 233–286

    Google Scholar 

  4. Hagen WR (1992) EPR spectroscopy of iron-sulfur proteins. In: Sykes AG, Cammack R (eds) Advances in inorganic chemistry: iron-sulfur proteins, vol 38. Academic Press, San Diego, CA, pp 165–222

    Google Scholar 

  5. Crouse BR, Yano T, Finnegan MG et al (1994) Properties of the iron-sulfur center in the 25-kilodalton subunit of the proton-translocating NADH-quinone oxidireductase of Paracoccus denitrificans. J Biol Chem 269:21030–21036

    CAS  PubMed  Google Scholar 

  6. Lindahl PA, Day EP, Kent TA et al (1985) Mössbauer, EPR, and magnetization studies of the Azotobacter vinelandii Fe protein. J Biol Chem 260:11160–11173

    CAS  PubMed  Google Scholar 

  7. Onate YA, Finnegan MG, Hales BJ et al (1993) Variable temperature magnetic circular dichroism studies of reduced nitrogenase iron proteins and [4Fe-4S]+ synthetic analog clusters. Biochim Biophys Acta 1164:113–123

    Article  CAS  Google Scholar 

  8. Angove HC, Yoo SJ, Burgess BK et al (1997) Mössbauer and EPR evidence for an all-ferrous Fe4S4 cluster with S = 4 in the Fe protein of nitrogenase. J Am Chem Soc 119:8730–8731

    Article  CAS  Google Scholar 

  9. Yoo SJ, Angove HC, Burgess BK et al (1998) Magnetic circular dichroism study of the all-ferrous [4Fe-4S] cluster of the Fe-protein of Azotobacter vinelandii nitrogenase. J Am Chem Soc 120:9704–9705

    Article  CAS  Google Scholar 

  10. Beinert H, Holm RH, Münck E (1997) Iron-sulfur clusters: Nature’s modular, multipurpose structures. Science 277:653–659

    Article  CAS  Google Scholar 

  11. Rupnik K, Lee CC, Hu Y et al (2011) [4Fe4S]2+ clusters exhibit ground-state paramagnetism. J Am Chem Soc 133:6871–6873

    Article  CAS  Google Scholar 

  12. Hagen WR, Wassink H, Eady RR et al (1987) Quantitative EPR of an S = 7/2 system in thionine-oxidized MoFe proteins of nitrogenase: a redefinition on the P-cluster concept. Eur J Chem 169:457–465

    Article  CAS  Google Scholar 

  13. Surerus KK, Hendrich MP, Christie PD et al (1992) Mössbauer and integer-spin EPR of the oxidized P-clusters of nitrogenase: Pox is a non-Kramers system with a nearly degenerate ground doublet. J Am Chem Soc 114:8579–8590

    Article  CAS  Google Scholar 

  14. Münck E, Rhodes H, Orme-Johnson WH et al (1975) The MoFe protein component from Azotobacter vinelandii. Biochim Biophys Acta 400:32–53

    Article  Google Scholar 

  15. Broach RB, Rupnik K, Hu Y et al (2004) VTVH-MCD spectroscopic study of the metal clusters in the ΔnifB and ΔnifH MoFe proteins of nitrogenase from Azotobacter vinelandii. Biochemistry 45:15039–15048

    Article  Google Scholar 

  16. Lindahl PA, Papaefthymiou V, Orme-Johnson WH et al (1988) Mössbauer studies of solid thionin-oxidized MoFe protein of nitrogenase. J Biol Chem 263:19412–19418

    CAS  PubMed  Google Scholar 

  17. Johnson MK, Thomson AJ, Robinson AE et al (1981) Characterization of the paramagnetic centres of the molybdenum-iron protein of nitrogenase from Klebsiella pneumoniae using low-temperature magnetic circular dichroism spectroscopy. Biochim Biophys Acta 671:61–70

    Article  CAS  Google Scholar 

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Correspondence to Brian J. Hales .

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Hales, B.J. (2019). Magnetic Circular Dichroism Spectroscopy of Metalloproteins. 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_14

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  • DOI: https://doi.org/10.1007/978-1-4939-8864-8_14

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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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