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Analyzing heme proteins using EPR techniques: the heme-pocket structure of ferric mouse neuroglobin

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Abstract

In this work, an electron paramagnetic resonance (EPR) strategy to study the heme-pocket structure of low-spin ferric heme proteins is optimized. Frozen solutions of ferric mouse neuroglobin (mNgb) are analyzed by means of electron spin echo envelope modulation and pulsed electron–nuclear double resonance techniques. The hyperfine and nuclear quadrupole couplings of the directly coordinating heme and histidine nitrogens are derived and are discussed in comparison with known data of other ferric porphyrin compounds. In combination with the hyperfine matrices of the imidazole protons, the 14N EPR parameters reveal structural information on the heme pocket of mNgb that is in agreement with previous X-ray diffraction data on neuroglobins.

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Abbreviations

CP:

Combination peak

CW:

Continuous wave

dq:

Double quantum

ENDOR:

Electron–nuclear double resonance

EPR:

Electron paramagnetic resonance

ESEEM:

Electron spin echo envelope modulation

Hb:

Hemoglobin

HYSCORE:

Hyperfine sublevel correlation

LS:

Low spin

Mb:

Myoglobin

4-MeIm:

4-Methyl imidazole

mNgb:

Mouse neuroglobin

mw:

Microwave

Ngb:

Neuroglobin

NGB:

Human neuroglobin

PPIX:

Protoporphyrin IX

rf:

Radio frequency

TPP:

Tetraphenylporphyrin

Tris:

Tris(hydroxymethyl)aminomethane

wt:

Wild type

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Acknowledgment

This work was supported by the Fund for Scientific Research-Flanders (FWO) Grant G.0468.03 (to S.V.D) and Grant QLRT-2001-01548 from the European Union. S.D. is a postdoctoral fellow of the FWO. E.V. is a research assistant of the FWO. S.V.D. wants to thank Martino Bolognesi (Universities of Milano and Genova) for interesting discussions on the X-ray data of Ngb proteins.

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Authors and Affiliations

  1. Department of Physics, University of Antwerp, Campus Drie Eiken, Universiteitsplein 1, 2610, Wilrijk, Belgium

    E. Vinck & S. Van Doorslaer

  2. Department of Biochemical Sciences, University of Antwerp, Antwerp, Belgium

    S. Dewilde & L. Moens

  3. Physical Chemistry Laboratory, ETH Hönggerberg, HCI, 8093, Zurich, Switzerland

    G. Mitrikas & A. Schweiger

Authors
  1. E. Vinck
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  2. S. Van Doorslaer
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  3. S. Dewilde
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  4. G. Mitrikas
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  5. A. Schweiger
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  6. L. Moens
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Corresponding author

Correspondence to S. Van Doorslaer.

Additional information

This paper is dedicated to our coauthor Prof. Arthur Schweiger, who passed away unexpectedly on 4 January 2006.

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Vinck, E., Van Doorslaer, S., Dewilde, S. et al. Analyzing heme proteins using EPR techniques: the heme-pocket structure of ferric mouse neuroglobin. J Biol Inorg Chem 11, 467–475 (2006). https://doi.org/10.1007/s00775-006-0100-2

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  • DOI: https://doi.org/10.1007/s00775-006-0100-2

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