Advertisement

Biomolecular NMR Assignments

, Volume 9, Issue 1, pp 197–200 | Cite as

Backbone assignments of the apo and Zn(II) protoporphyrin IX-bound states of the soluble form of rat heme oxygenase-1

  • Erisa Harada
  • Masakazu Sugishima
  • Jiro Harada
  • Masato Noguchi
  • Keiichi Fukuyama
  • Kenji SugaseEmail author
Article

Abstract

In nature, heme is a prosthetic group that is universally used as a cofactor for heme proteins. It is necessary for the execution of fundamental biological processes including electron transfer, oxidation and metabolism. However, free heme is toxic to cells, because of its capability to enhance oxidative stress, hence its cellular concentration is strictly regulated through multiple mechanisms. Heme oxygenase (HO) serves as an irreplaceable member in the heme degradation system. It is a ubiquitous protein, existing in many species including mammals, higher plants, and interestingly, certain pathogenic bacteria. In the HO reaction, HO catalyzes oxidative cleavage of heme to generate biliverdin and release carbon monoxide and ferrous iron. Because of the beneficial effects of these heme catabolism products, HO plays a key role in iron homeostasis and in defense mechanism against oxidative stress. HO is composed of an N-terminal structured region and a C-terminal membrane-bound region. Furthermore, the soluble form of HO, which is obtainable by excision of the membrane-bound region, retains its catalytic activity. Here, we present the backbone resonance assignments of the soluble form (residues 1–232) of HO-1 in the free and Zn(II) protoporphyrin IX (ZnPP)-bound states, and analyzed the structural differences between the states. ZnPP is a potent enzyme inhibitor, and the ZnPP-bound structure of HO-1 mimics the heme-bound structure. These assignments provide the structural basis for a detailed investigation of the HO-1 function.

Keywords

Heme oxygenase NMR resonance assignments Cellular homeostasis Heme protein 

Notes

Acknowledgments

We thank Naohiro Kobayashi for assistance in NMR chemical shift assignments. This work was supported by Grants-in-Aid for Scientific Research for KS, MS, JH, MN, and KF from the Ministry of Education, Culture, Sports and Technology of Japan.

Supplementary material

12104_2014_9573_MOESM1_ESM.pdf (458 kb)
Supplementary material 1 (PDF 458 kb)

References

  1. Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293. doi: 10.1007/BF00197809 CrossRefGoogle Scholar
  2. Khan AA, Quigley JG (2011) Control of intracellular heme levels: heme transporters and heme oxygenases. Biochem Biophys Acta 1813:668–682. doi: 10.1016/j.bbamcr.2011.01.008 CrossRefGoogle Scholar
  3. Kobayashi N, Iwahara J, Koshiba S, Tomizawa T, Tochio N, Güntert P, Kigawa T, Yokoyama S (2007) KUJIRA, a package of integrated modules for systematic and interactive analysis of NMR data directed to high-throughput NMR structure studies. J Biomol NMR 39:31–52. doi: 10.1007/s10858-007-9175-5 CrossRefGoogle Scholar
  4. Li Y, Syvitski RT, Auclair K, Ortiz de Montellano PR, La Mar GN (2002) Solution NMR characterization of an unusual distal H-bond network in the active site of the cyanide-inhibited, human heme oxygenase complex of the symmetric substrate, 2,4-dimethyldeuterohemin. J Biol Chem 277:33018–33031. doi: 10.1074/jbc.M204216200
  5. Li Y, Syvitski RT, Auclair K, Ortiz de Montellano PR, La Mar GN (2004) 1H NMR investigation of the solution structure of substrate-free human heme oxygenase: comparison to the cyanide-inhibited, substrate-bound complex. J Biol Chem 279:10195–10205. doi: 10.1074/jbc.M308379200
  6. Omata Y, Asada S, Sakamoto H, Fukuyama K, Noguchi M (1998) Crystallization and preliminary X-ray diffraction studies on the water soluble form of rat heme oxygenase-1 in complex with heme. Acta Crystallogr D54:1017–1019Google Scholar
  7. Schuller DJ, Wilks A, Ortiz  de Montellano PR, Poulos TL (1999) Crystal structure of human heme oxygenase-1. Nat Struct Biol 6:860–867. doi: 10.1038/12319
  8. Sugishima M, Omata Y, Kakuta Y, Sakamoto H, Noguchi M, Fukuyama K (2000) Crystal structure of rat heme oxygenase-1 in complex with heme. FEBS Lett 471:61–66CrossRefGoogle Scholar
  9. Sugishima M, Sakamoto H, Kakuta Y, Omata Y, Hayashi S, Noguchi M, Fukuyama K (2002) Crystal structure of rat apo-heme oxygenase-1 (HO-1): mechanism of heme binding in HO-1 inferred from structural comparison of the apo and heme complex forms. Biochemistry 41:7293–7300CrossRefGoogle Scholar
  10. Verma A, Hirsch DJ, Glatt CE, Ronnett GV, Snyder SH (1993) Carbon monoxide: a putative neural messenger. Science 259:381–384CrossRefADSGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Erisa Harada
    • 1
  • Masakazu Sugishima
    • 2
  • Jiro Harada
    • 2
  • Masato Noguchi
    • 2
    • 4
  • Keiichi Fukuyama
    • 3
    • 5
  • Kenji Sugase
    • 1
    Email author
  1. 1.Bioorganic Research InstituteSuntory Foundation for Life SciencesMishimaJapan
  2. 2.Kurume University School of MedicineKurumeJapan
  3. 3.Graduate School of ScienceOsaka UniversityToyonakaJapan
  4. 4.Faculty of Fukuoka Medical TechnologyTeikyo UniversityOmutaJapan
  5. 5.Graduate School of EngineeringOsaka UniversitySuitaJapan

Personalised recommendations