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Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem

Nature volume 435, pages 697–701 (2005)Cite this article

Abstract

The synthesis of haemoglobin A (HbA) is exquisitely coordinated during erythrocyte development to prevent damaging effects from individual α- and β-subunits1,2. The α-haemoglobin-stabilizing protein (AHSP) binds α-haemoglobin (αHb), inhibits the ability of αHb to generate reactive oxygen species and prevents its precipitation on exposure to oxidant stress3,4,5. The structure of AHSP bound to ferrous αHb is thought to represent a transitional complex through which αHb is converted to a non-reactive, hexacoordinate ferric form5. Here we report the crystal structure of this ferric αHb–AHSP complex at 2.4 Å resolution. Our findings reveal a striking bis-histidyl configuration in which both the proximal and the distal histidines coordinate the haem iron atom. To attain this unusual conformation, segments of αHb undergo drastic structural rearrangements, including the repositioning of several α-helices. Moreover, conversion to the ferric bis-histidine configuration strongly and specifically inhibits redox chemistry catalysis and haem loss from αHb. The observed structural changes, which impair the chemical reactivity of haem iron, explain how AHSP stabilizes αHb and prevents its damaging effects in cells.

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Figure 1: Structure of the oxidized αHb bound to AHSP.
Figure 2: AHSP-bound oxidized αHb undergoes structural rearrangements.
Figure 3: Protective effects of the bis-histidyl configuration for Fe  iii -αHb.
Figure 4: Mechanisms of AHSP-mediated stabilization of αHb.

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Acknowledgements

We thank M. Becker and A. Sexana at BNL NSLS beamlines for help. This work was supported by grants from the National Institutes of Health (to Y.S. and M.J.W.), the American Heart Association (to A.J.G.) and the Cooley's Anemia Foundation (S.Z.).Author Contributions L.F., S.Z. and L.G. contributed equally to this work. L.F. crystallized the oxidized α-haemoglobin bound to AHSP. L.F. and L.G. solved the structure. S.Z. and L.F. performed the biochemical experiments in Fig. 3. Y.S. and A.J.G. wrote the paper. All authors discussed the results and commented on the manuscript.

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Authors and Affiliations

  1. Department of Molecular Biology, Lewis Thomas Laboratory, Princeton University, Princeton, New Jersey, 08544, USA

    Liang Feng, Lichuan Gu & Yigong Shi

  2. The Children's Hospital of Philadelphia and the University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA

    Suiping Zhou, Mitchell J. Weiss & Andrew J. Gow

  3. School of Molecular and Microbial Biosciences, University of Sydney, NSW, 2006, Australia

    David A. Gell & Joel P. Mackay

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Correspondence to Andrew J. Gow or Yigong Shi.

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The atomic coordinates of the oxidized αHb–AHSP complex have been deposited in the Protein Data Bank with the accession number 1Z8U. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure S1

Structural changes in the haeme-binding region of αHb (JPG 134 kb)

Supplementary Figure S2

Quantification of the change in haeme species, after the addition of hydrogen peroxide to the ferrous αHb complex (panel a) and the ferric Hb complex (panel b). (JPG 45 kb)

Supplementary Table S1

Summary of crystallographic analysis (DOC 22 kb)

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Feng, L., Zhou, S., Gu, L. et al. Structure of oxidized α-haemoglobin bound to AHSP reveals a protective mechanism for haem. Nature 435, 697–701 (2005). https://doi.org/10.1038/nature03609

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