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PMID: 18446211 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

NO dioxygenase activity in hemoglobins is ubiquitous in vitro, but limited by reduction in vivo.

PloS one ·Vol. 3 ·No. 4 ·2008-04-30 ·Pages e2039

Smagghe BJ, Trent JT, Hargrove MS

Abstract

Genomics has produced hundreds of new hemoglobin sequences with examples in nearly every living organism. Structural and biochemical characterizations of many recombinant proteins reveal reactions, like oxygen binding and NO dioxygenation, that appear general to the hemoglobin superfamily regardless of whether they are related to physiological function. Despite considerable attention to "hexacoordinate" hemoglobins, which are found in nearly every plant and animal, no clear physiological role(s) has been assigned to them in any species. One popular and relevant hypothesis for their function is protection against NO. Here we have tested a comprehensive representation of hexacoordinate hemoglobins from plants (rice hemoglobin), animals (neuroglobin and cytoglobin), and bacteria (Synechocystis hemoglobin) for their abilities to scavenge NO compared to myoglobin. Our experiments include in vitro comparisons of NO dioxygenation, ferric NO binding, NO-induced reduction, NO scavenging with an artificial reduction system, and the ability to substitute for a known NO scavenger (flavohemoglobin) in E. coli. We conclude that none of these tests reveal any distinguishing predisposition toward a role in NO scavenging for the hxHbs, but that any hemoglobin could likely serve this role in the presence of a mechanism for heme iron re-reduction. Hence, future research to test the role of Hbs in NO scavenging would benefit more from the identification of cognate reductases than from in vitro analysis of NO and O(2) binding.

MeSH Terms
Animals Catalysis Escherichia coli Ferredoxins/metabolism Free Radical Scavengers/metabolism Heme/metabolism Hemoglobins/metabolism Horses Humans Iron/metabolism Kinetics Mutation Myoglobin/metabolism Nitric Oxide/metabolism Oryza/metabolism Oxidation-Reduction Oxidoreductases/metabolism Oxygen/metabolism Oxygenases/metabolism Spectrum Analysis Synechocystis/metabolism
Chemicals
Ferredoxins Free Radical Scavengers Hemoglobins Myoglobin Nitric Oxide Heme Iron Oxidoreductases Oxygenases nitric oxide dioxygenase Oxygen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Smagghe Benoit J
Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, Iowa, United States of America.
Trent James T
Hargrove Mark S
References (83)
83 references, click to expand
  1. Cytoglobin: a novel globin type ubiquitously expressed in vertebrate tissues.
    Mol Biol Evol. 2002 Apr;19(4):416-21 PMID: 11919282
  2. Mechanistic studies of the oxygen-mediated oxidation of nitrosylhemoglobin.
    Biochemistry. 2005 Apr 26;44(16):6223-31 PMID: 15835910
  3. Nitric oxide dioxygenase function and mechanism of flavohemoglobin, hemoglobin, myoglobin and their associated reductases.
    J Inorg Biochem. 2005 Jan;99(1):247-66 PMID: 15598505
  4. Transcriptional responses of Escherichia coli to S-nitrosoglutathione under defined chemostat conditions reveal major changes in methionine biosynthesis.
    J Biol Chem. 2005 Mar 18;280(11):10065-72 PMID: 15647275
  5. Crystal structure of cytoglobin: the fourth globin type discovered in man displays heme hexa-coordination.
    J Mol Biol. 2004 Feb 27;336(4):917-27 PMID: 15095869
  6. Nitric oxide scavenging and detoxification by the Mycobacterium tuberculosis haemoglobin, HbN in Escherichia coli.
    Mol Microbiol. 2002 Sep;45(5):1303-14 PMID: 12207698
  7. Arabidopsis nonsymbiotic hemoglobin AHb1 modulates nitric oxide bioactivity.
    Plant Cell. 2004 Oct;16(10):2785-94 PMID: 15367716
  8. Mechanism of NO-induced oxidation of myoglobin and hemoglobin.
    Biochemistry. 1996 Jun 4;35(22):6976-83 PMID: 8679521
  9. Protection from nitrosative stress by yeast flavohemoglobin.
    Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4672-6 PMID: 10758168
  10. Hemoglobins dioxygenate nitric oxide with high fidelity.
    J Inorg Biochem. 2006 Apr;100(4):542-50 PMID: 16439024
  11. Bis-histidyl hexacoordination in hemoglobins facilitates heme reduction kinetics.
    J Am Chem Soc. 2004 Sep 29;126(38):11930-5 PMID: 15382928
  12. Three distinct Arabidopsis hemoglobins exhibit peroxidase-like activity and differentially mediate nitrite-dependent protein nitration.
    FEBS Lett. 2004 Aug 13;572(1-3):27-32 PMID: 15304319
  13. The crystal structure of Synechocystis hemoglobin with a covalent heme linkage.
    J Biol Chem. 2004 Apr 16;279(16):16535-42 PMID: 14736872
  14. Neuroglobin is up-regulated by and protects neurons from hypoxic-ischemic injury.
    Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15306-11 PMID: 11742077
  15. Flavohemoglobin requires microaerophilic conditions for nitrosative protection of Staphylococcus aureus.
    FEBS Lett. 2006 Mar 20;580(7):1817-21 PMID: 16516202
  16. Flavohemoglobin denitrosylase catalyzes the reaction of a nitroxyl equivalent with molecular oxygen.
    Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10108-12 PMID: 11517313
  17. The reaction of neuroglobin with potential redox protein partners cytochrome b5 and cytochrome c.
    FEBS Lett. 2006 Sep 4;580(20):4884-8 PMID: 16914148
  18. Nitric oxide dioxygenase: an enzymic function for flavohemoglobin.
    Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10378-83 PMID: 9724711
  19. Nitric-oxide dioxygenase activity and function of flavohemoglobins. sensitivity to nitric oxide and carbon monoxide inhibition.
    J Biol Chem. 2000 Oct 13;275(41):31581-7 PMID: 10922365
  20. The X-ray structure of ferric Escherichia coli flavohemoglobin reveals an unexpected geometry of the distal heme pocket.
    J Biol Chem. 2002 Jun 28;277(26):23725-32 PMID: 11964402
  21. NADH-dependent methemoglobin reductase from the obligate aerobe Vitreoscilla: improved method of purification and reexamination of prosthetic groups.
    Arch Biochem Biophys. 1992 Jan;292(1):29-33 PMID: 1309298
  22. Modulation of nitric oxide bioactivity by plant haemoglobins.
    J Exp Bot. 2006;57(3):479-88 PMID: 16377734
  23. Plants, humans and hemoglobins.
    Trends Plant Sci. 2003 Aug;8(8):387-93 PMID: 12927972
  24. Truncated hemoglobin HbN protects Mycobacterium bovis from nitric oxide.
    Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):5902-7 PMID: 11959913
  25. A ubiquitously expressed human hexacoordinate hemoglobin.
    J Biol Chem. 2002 May 31;277(22):19538-45 PMID: 11893755
  26. Truncated hemoglobin from the cyanobacterium Synechococcus sp. PCC 7002: evidence for hexacoordination and covalent adduct formation in the ferric recombinant protein.
    Biochemistry. 2002 Jun 4;41(22):6902-10 PMID: 12033922
  27. Bacterial hemoglobins and flavohemoglobins for alleviation of nitrosative stress in Escherichia coli.
    Appl Environ Microbiol. 2002 Oct;68(10):4835-40 PMID: 12324328
  28. A vertebrate globin expressed in the brain.
    Nature. 2000 Sep 28;407(6803):520-3 PMID: 11029004
  29. Neuroglobin protects the brain from experimental stroke in vivo.
    Proc Natl Acad Sci U S A. 2003 Mar 18;100(6):3497-500 PMID: 12621155
  30. Slow ligand binding kinetics dominate ferrous hexacoordinate hemoglobin reactivities and reveal differences between plants and other species.
    Biochemistry. 2006 Jan 17;45(2):561-70 PMID: 16401085
  31. Crystal structure of a nonsymbiotic plant hemoglobin.
    Structure. 2000 Sep 15;8(9):1005-14 PMID: 10986467
  32. Nitric oxide binding properties of neuroglobin. A characterization by EPR and flash photolysis.
    J Biol Chem. 2003 Feb 14;278(7):4919-25 PMID: 12480932
  33. Kinetic and mechanistic studies of the NO*-mediated oxidation of oxymyoglobin and oxyhemoglobin.
    Biochemistry. 2001 Mar 20;40(11):3385-95 PMID: 11258960
  34. Characterization of a stellate cell activation-associated protein (STAP) with peroxidase activity found in rat hepatic stellate cells.
    J Biol Chem. 2001 Jul 6;276(27):25318-23 PMID: 11320098
  35. S-nitrosohaemoglobin: a dynamic activity of blood involved in vascular control.
    Nature. 1996 Mar 21;380(6571):221-6 PMID: 8637569
  36. Ascaris haemoglobin is a nitric oxide-activated 'deoxygenase'.
    Nature. 1999 Sep 30;401(6752):497-502 PMID: 10519555
  37. Nitric oxide sensitivity of the aconitases.
    J Biol Chem. 1997 Oct 3;272(40):25071-6 PMID: 9312115
  38. Capillary zone electrophoretic detection of biological thiols and their S-nitrosated derivatives.
    Anal Chem. 1992 Apr 1;64(7):779-85 PMID: 1524223
  39. A brief history of hemoglobins: plant, animal, protist, and bacteria.
    Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5675-9 PMID: 8650150
  40. Human brain neuroglobin structure reveals a distinct mode of controlling oxygen affinity.
    Structure. 2003 Sep;11(9):1087-95 PMID: 12962627
  41. Biochemical characterization and ligand binding properties of neuroglobin, a novel member of the globin family.
    J Biol Chem. 2001 Oct 19;276(42):38949-55 PMID: 11473128
  42. An electrostatic model for the frequency shifts in the carbonmonoxy stretching band of myoglobin: correlation of hydrogen bonding and the stark tuning rate.
    J Am Chem Soc. 2002 Nov 6;124(44):13271-81 PMID: 12405856
  43. Cellular mechanism of oxygen sensing.
    Annu Rev Physiol. 2001;63:259-87 PMID: 11181957
  44. Mechanisms of ferriheme reduction by nitric oxide: nitrite and general base catalysis.
    Inorg Chem. 2004 Aug 23;43(17):5393-402 PMID: 15310219
  45. Nitric oxide moves myoglobin centre stage.
    Trends Biochem Sci. 2001 Apr;26(4):209-10 PMID: 11295538
  46. Neuroglobin, nitric oxide, and oxygen: functional pathways and conformational changes.
    Proc Natl Acad Sci U S A. 2005 Jun 14;102(24):8483-8 PMID: 15932948
  47. Nitric oxide scavenging by barley hemoglobin is facilitated by a monodehydroascorbate reductase-mediated ascorbate reduction of methemoglobin.
    Planta. 2006 Apr;223(5):1033-40 PMID: 16341544
  48. Assay and characterization of the NO dioxygenase activity of flavohemoglobins.
    Methods Enzymol. 2008;436:217-37 PMID: 18237635
  49. INTERRELATIONSHIP BETWEEN STRUCTURE AND FUNCTION IN HEMOGLOBIN AND MYOGLOBIN.
    Physiol Rev. 1965 Jan;45:123-70 PMID: 14257159
  50. A novel mechanism for upregulation of the Escherichia coli K-12 hmp (flavohaemoglobin) gene by the 'NO releaser', S-nitrosoglutathione: nitrosation of homocysteine and modulation of MetR binding to the glyA-hmp intergenic region.
    Mol Microbiol. 1998 Aug;29(4):1101-12 PMID: 9767577
  51. Flavohemoglobin detoxifies nitric oxide in aerobic, but not anaerobic, Escherichia coli. Evidence for a novel inducible anaerobic nitric oxide-scavenging activity.
    J Biol Chem. 2002 Mar 8;277(10):8166-71 PMID: 11751864
  52. Yeast flavohemoglobin is an ancient protein related to globins and a reductase family.
    Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5015-9 PMID: 1594608
  53. Neuroglobins from the zebrafish Danio rerio and the pufferfish Tetraodon nigroviridis.
    Biochem Biophys Res Commun. 2001 Sep 21;287(2):418-21 PMID: 11554744
  54. A flash photolysis method to characterize hexacoordinate hemoglobin kinetics.
    Biophys J. 2000 Nov;79(5):2733-8 PMID: 11053146
  55. Mechanism of nitric oxide release from S-nitrosothiols.
    J Biol Chem. 1996 Aug 2;271(31):18596-603 PMID: 8702510
  56. Class-1 hemoglobins, nitrate and NO levels in anoxic maize cell-suspension cultures.
    Planta. 2004 May;219(1):66-72 PMID: 14963709
  57. The role of glutathione in the transport and catabolism of nitric oxide.
    FEBS Lett. 1996 Mar 18;382(3):223-8 PMID: 8605974
  58. An enzymic reduction system for metmyoglobin and methemoglobin, and its application to functional studies of oxygen carriers.
    Biochim Biophys Acta. 1973 Jun 15;310(2):309-16 PMID: 4146292
  59. Truncated hemoglobins: a new family of hemoglobins widely distributed in bacteria, unicellular eukaryotes, and plants.
    J Biol Chem. 2002 Jan 11;277(2):871-4 PMID: 11696555
  60. The flavohemoglobin of Escherichia coli confers resistance to a nitrosating agent, a "Nitric oxide Releaser," and paraquat and is essential for transcriptional responses to oxidative stress.
    J Biol Chem. 1999 Jan 8;274(2):748-54 PMID: 9873011
  61. Functional adaptation and its molecular basis in vertebrate hemoglobins, neuroglobins and cytoglobins.
    Respir Physiol Neurobiol. 2004 Dec 15;144(2-3):141-59 PMID: 15556098
  62. Mechanisms of reductive nitrosylation in iron and copper models relevant to biological systems.
    Chem Rev. 2005 Jun;105(6):2439-55 PMID: 15941218
  63. Steady-state and transient kinetics of Escherichia coli nitric-oxide dioxygenase (flavohemoglobin). The B10 tyrosine hydroxyl is essential for dioxygen binding and catalysis.
    J Biol Chem. 2000 Apr 28;275(17):12581-9 PMID: 10777548
  64. Rate of reaction with nitric oxide determines the hypertensive effect of cell-free hemoglobin.
    Nat Biotechnol. 1998 Jul;16(7):672-6 PMID: 9661203
  65. Nitric oxide in brain: diffusion, targets and concentration dynamics in hippocampal subregions.
    Mol Aspects Med. 2004 Feb-Apr;25(1-2):75-89 PMID: 15051318
  66. Myoglobin: A scavenger of bioactive NO.
    Proc Natl Acad Sci U S A. 2001 Jan 16;98(2):735-40 PMID: 11136228
  67. Neuroglobin and cytoglobin in search of their role in the vertebrate globin family.
    J Inorg Biochem. 2005 Jan;99(1):110-9 PMID: 15598495
  68. Nitrate, NO and haemoglobin in plant adaptation to hypoxia: an alternative to classic fermentation pathways.
    J Exp Bot. 2004 Dec;55(408):2473-82 PMID: 15448180
  69. A model for ligand binding to hexacoordinate hemoglobins.
    Biochemistry. 2001 May 22;40(20):6155-63 PMID: 11352753
  70. No scavenging and the hypertensive effect of hemoglobin-based blood substitutes.
    Free Radic Biol Med. 2004 Mar 15;36(6):685-97 PMID: 14990349
  71. Analysis of the neuroprotective effects of various nitric oxide donor compounds in murine mixed cortical cell culture.
    J Neurochem. 1999 May;72(5):1843-52 PMID: 10217260
  72. Nonenzymatic reduction and oxidation of myoglobin and hemoglobin by nicotinamide adenine dinucleotides and flavins.
    J Biol Chem. 1969 Dec 25;244(24):6702-6 PMID: 4391279
  73. Influence of the protein matrix on intramolecular histidine ligation in ferric and ferrous hexacoordinate hemoglobins.
    Proteins. 2007 Jan 1;66(1):172-82 PMID: 17044063
  74. Imidazole antibiotics inhibit the nitric oxide dioxygenase function of microbial flavohemoglobin.
    Antimicrob Agents Chemother. 2005 May;49(5):1837-43 PMID: 15855504
  75. Reactions of peroxynitrite with globin proteins and their possible physiological role.
    Comp Biochem Physiol A Mol Integr Physiol. 2005 Oct;142(2):124-9 PMID: 16055362
  76. Ligand binding and hexacoordination in synechocystis hemoglobin.
    J Biol Chem. 2001 Sep 14;276(37):34714-21 PMID: 11438545
  77. Myoglobin-facilitated diffusion of oxygen.
    J Gen Physiol. 1965 Sep;49(1):Suppl:57-74 PMID: 5859926
  78. Flavohemoglobin Hmp, but not its individual domains, confers protection from respiratory inhibition by nitric oxide in Escherichia coli.
    J Biol Chem. 2003 Sep 12;278(37):34975-82 PMID: 12826671
  79. Chimeric Vitreoscilla hemoglobin (VHb) carrying a flavoreductase domain relieves nitrosative stress in Escherichia coli: new insight into the functional role of VHb.
    Appl Environ Microbiol. 2002 Jan;68(1):152-60 PMID: 11772621
  80. How does the eye breathe? Evidence for neuroglobin-mediated oxygen supply in the mammalian retina.
    J Biol Chem. 2003 Jan 17;278(3):1932-5 PMID: 12409290
  81. Neuroglobin and cytoglobin as potential enzyme or substrate.
    Gene. 2007 Aug 15;398(1-2):103-13 PMID: 17555889
  82. Reactivity studies of the Fe(III) and Fe(II)NO forms of human neuroglobin reveal a potential role against oxidative stress.
    J Biol Chem. 2004 May 28;279(22):22841-7 PMID: 15020597
  83. Human neuroglobin, a hexacoordinate hemoglobin that reversibly binds oxygen.
    J Biol Chem. 2001 Aug 10;276(32):30106-10 PMID: 11429401
Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0002039
Published
2008-04-30
Epub
2008-00-30
Pages
e2039
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC2323109
Subset
IM
Grants
NIGMS NIH HHS · R01 GM065948 · United States
NIGMS NIH HHS · R01-GM065948 · United States
External Links
PubMed source DOI: 10.1371/journal.pone.0002039 PMC Full Text View journal details
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