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Transition Metals in Biochemistry

Time-resolved FTIR study of CO recombination with horseradish peroxidase

Amandine Maréchal, W. John Ingledew, Peter R. Rich
Biochemical Society Transactions Dec 01, 2008, 36 (6) 1165-1168; DOI: 10.1042/BST0361165
Amandine Maréchal
Glynn Laboratory of Bioenergetics, Institute of Structural and Molecular Biology, University College London, Gower Street, London WC1 6BT, U.K.
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W. John Ingledew
School of Biology, University of St. Andrews, St. Andrews KY16 9ST, U.K.
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Peter R. Rich
Glynn Laboratory of Bioenergetics, Institute of Structural and Molecular Biology, University College London, Gower Street, London WC1 6BT, U.K.
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Abstract

Vibrational changes associated with CO recombination to ferrous horseradish peroxidase were investigated by rapid-scan FTIR (Fourier-transform IR) spectroscopy in the 1200–2200 cm−1 range. At pH 6.0, two conformers of bound CO are present that appear as negative bands at 1905 and 1934 cm−1 in photolysis spectra. Their recombination rate constants are identical, confirming that they arise from two substates of bound CO that are in rapid thermal equilibrium, rather than from heterogeneous protein sites. A smaller positive band at 2134 cm−1 also appears on photolysis and decays with the same rate constant, indicative of an intraprotein geminate site involved in recombination or, possibly, a weak-affinity surface CO-binding site. Other signals arising from protein and haem in the 1700–1200 cm−1 range can also be time-resolved with similar kinetics.

  • carbon monoxide
  • Fourier-transform infrared spectroscopy (FTIR spectroscopy)
  • horseradish peroxidase
  • rapid-scan
  • time-resolved spectroscopy

Footnotes

  • Transition Metals in Biochemistry: A joint Biochemical Society meeting with the Inorganic Biochemistry Discussion Group to honour Professor Andrew Thomson FRS, held at University of East Anglia, Norwich, U.K., 24–26 June 2008. Organized and Edited by Steve Chapman (Edinburgh, U.K.), David Richardson (University of East Anglia, U.K.) and Nick Watmough (University of East Anglia, U.K.).

Abbreviations: FTIR, Fourier-transform IR; HRPC, horseradish peroxidase isoenzyme C

  • © The Authors Journal compilation © 2008 Biochemical Society
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December 2008

Volume: 36 Issue: 6

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Time-resolved FTIR study of CO recombination with horseradish peroxidase
Amandine Maréchal, W. John Ingledew, Peter R. Rich
Biochemical Society Transactions Dec 2008, 36 (6) 1165-1168; DOI: 10.1042/BST0361165
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Time-resolved FTIR study of CO recombination with horseradish peroxidase
Amandine Maréchal, W. John Ingledew, Peter R. Rich
Biochemical Society Transactions Dec 2008, 36 (6) 1165-1168; DOI: 10.1042/BST0361165

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  • Article
    • Abstract
    • Introduction
    • Time-resolved FTIR spectroscopy
    • Kinetic analyses
    • Kinetics of CO rebinding to ferrous HRPC
    • Vibrational modes associated with protein and haem
    • Conclusions
    • Acknowledgments
    • Footnotes
    • References
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Keywords

carbon monoxide
Fourier-transform infrared spectroscopy (FTIR spectroscopy)
horseradish peroxidase
rapid-scan
time-resolved spectroscopy

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