Advertisement

T-state Stabilization of Hemoglobin by Nitric Oxide to Form α-Nitrosyl Heme Causes Constitutive Release of ATP from Human Erythrocytes

  • Tomotaka Akatsu
  • Kosuke Tsukada
  • Takako Hishiki
  • Kazuhiro Suga-numa
  • Minoru Tanabe
  • Motohide Shimazu
  • Yuko Kitagawa
  • Ayako Yachie-Kinoshita
  • Makoto Suematsu
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 662)

Abstract

Upon hypoxia, erythrocytes utilize hemoglobin (Hb) to trigger acti-vation of glycolysis through its interaction with band 3. This process contributes to maintenance of ATP, a portion of which is released extracellularly to trigger endothelium-dependent vasorelaxation. However, whether the ATP release results either from metabolic activation of the cells secondarily or from direct regulation of the gating through Hb allostery remains unknown. This study aimed to examine if stabilization of T-state Hb could induce steady-state and hypoxia-induced alterations in glycolysis and the ATP release from erythrocytes. Treatment of deoxygenated erythrocytes with a nitric oxide (NO) donor generated α-NO Hb that is stabilized T-state allostery. Under these circumstances, the release of ATP was significantly elevated even under normoxia and not further enhanced upon hypoxia. These events did not coincide with activation of glycolysis of the cells, so far as judged by the fact that intracellular ATP was significantly decreased by the NO treatment. Collectively, the present study suggests that hypoxia-induced ATP release is triggered through mechanisms involving R-T transition of Hb, and the gating process might occur irrespective of hypoxia-responsive regulation of glycolysis.

Keywords

Nitric Oxide Electron Spin Resonance Metabolome Analysis Hypoxic Vasodilation Hypoxic Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported by Grant-in-Aid for Creative Scientific Research 17GS0419 from MEXT Japan.

References

  1. 1.
    S. Locovei, L. Bao, G. Dahl. Pannexin 1 in erythrocytes: function without a gap. Proc. Natl. Acad. Sci. U. S. A. 103, 7655–7659 (2006).PubMedCrossRefGoogle Scholar
  2. 2.
    M. L. Ellsworth, T. Forrester, C. G. Ellis, H. H. Dietrich. The erythrocyte as a regulator of vascular tone. Am. J. Physiol. Heart Circ. Physiol. 269, 2155–2161 (1995).Google Scholar
  3. 3.
    K. Yamamoto, T. Sokabe, T. Matsumoto, K. Yoshimura, M. Shibata, N. Ohura, T. Fukuda, T. Sato, K. Sekine, S. Kato, M. Isshiki, T. Fujita, M. Kobayashi, K. Kawamura, H. Masuda, A. Kamiya, J. Ando. Impaired flow-dependent control of vascular tone and remodeling in P2X4-deficient mice. Nat. Med. 12, 133–137 (2006).PubMedCrossRefGoogle Scholar
  4. 4.
    J. E. Jagger, R. M. Bateman, M. L. Ellsworth, C. G. Ellis. Role of erythrocyte in regulating local O2 delivery mediated by hemoglobin oxygenation. Am. J. Physiol. Heart Circ. Physiol. 280, 2833–2839 (2001).Google Scholar
  5. 5.
    J. Gonzalez-Alonso, D. B. Olsen, B. Saltin. Erythrocyte and the regulation of human skeletal muscle blood flow and oxygen delivery: role of circulating ATP. Circ. Res. 91, 1046–1055 (2002).PubMedCrossRefGoogle Scholar
  6. 6.
    G. R. Bergfeld, T. Forrester. Release of ATP from human erythrocytes in response to a brief period of hypoxia and hypercapnia. Cardiovasc. Res. 26, 40–47 (1992).PubMedCrossRefGoogle Scholar
  7. 7.
    A. Kinoshita, K. Tsukada, T. Soga, T. Hishiki, Y. Ueno, Y. Nakayama, M. Tomita, M. Suematsu. Roles of hemoglobin allostery in hypoxia-induced metabolic alterations in erythrocytes: simulation and its verification by metabolome analysis. J. Biol. Chem. 282, 10731–10741 (2007).PubMedCrossRefGoogle Scholar
  8. 8.
    K. Suganuma, K. Tsukada, M. Kashiba, A. Tsuneshige, T. Furukawa, T. Kubota, N. Goda, M. Kitajima, T. Yonetani, M. Suematsu. Erythrocytes with T-state-stabilized hemoglobin as a therapeutic tool for postischemic liver dysfunction. Antioxid. Redox. Signal. 8, 1847–1855 (2006).PubMedCrossRefGoogle Scholar
  9. 9.
    M. Palacios-Callender, V. Hollis, M. Mitchison, N. Frakich, D. Unitt, S. Moncada. Cytochrome c oxidase regulates endogenous nitric oxide availability in respiring cells: A possible explanation for hypoxic vasodilatation. Proc. Natl. Acad. Sci. U. S. A. 104, 18508–18513 (2007).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Tomotaka Akatsu
    • 1
  • Kosuke Tsukada
    • 2
  • Takako Hishiki
    • 2
  • Kazuhiro Suga-numa
    • 1
  • Minoru Tanabe
    • 1
  • Motohide Shimazu
    • 1
  • Yuko Kitagawa
    • 3
  • Ayako Yachie-Kinoshita
    • 4
  • Makoto Suematsu
    • 2
  1. 1.Departments of SurgerySchool of Medicine, Keio UniversityTokyoJapan
  2. 2.Department of Biochemistry and Integrative Medical BiologySchool of Medicine, Keio UniversityTokyoJapan
  3. 3.Departments of SurgerySchool of Medicine, Keio UniversityTokyoJapan
  4. 4.Departments of Biochemistry and Integrative Medical BiologyKeio UniversityTokyoJapan

Personalised recommendations