Chemical reactivity of Synechococcus sp. PCC 7002 and Synechocystis sp. PCC 6803 hemoglobins: covalent heme attachment and bishistidine coordination

  • Henry J. Nothnagel
  • Matthew R. Preimesberger
  • Matthew P. Pond
  • Benjamin Y. Winer
  • Emily M. Adney
  • Juliette T. J. Lecomte
Original Paper

Abstract

In the absence of an exogenous ligand, the hemoglobins from the cyanobacteria Synechocystis sp. PCC 6803 and Synechococcus sp. PCC 7002 coordinate the heme group with two axial histidines (His46 and His70). These globins also form a covalent linkage between the heme 2-vinyl substituent and His117. The in vitro mechanism of heme attachment to His117 was examined with a combination of site-directed mutagenesis, NMR spectroscopy, and optical spectroscopy. The results supported an electrophilic addition with vinyl protonation being the rate-determining step. Replacement of His117 with a cysteine demonstrated that the reaction could occur with an alternative nucleophile. His46 (distal histidine) was implicated in the specificity of the reaction for the 2-vinyl group as well as protection of the protein from oxidative damage caused by exposure to exogenous H2O2.

Keywords

Truncated hemoglobin Heme posttranslational modification Hybrid b/c heme Reactive oxygen species Cytochrome c 

Abbreviations

DT

Dithionite

DTT

Dithiothreitol

GODCAT

Glucose oxidase/d-glucose/catalase

HMQC

Heteronuclear multiple quantum coherence

HSQC

Heteronuclear single quantum coherence

rGlbN

Recombinant GlbN

rGlbN-A

Recombinant GlbN with covalently attached heme

rGlbN-R

Recombinant GlbN prepared by reconstitution of the apoprotein with iron(III) protoporphyrin IX

RNS

Reactive nitrogen species

ROS

Reactive oxygen species

Synechococcus 7002

Synechococcus sp. PCC 7002

Synechocystis 6803

Synechocystis sp. PCC 6803

Supplementary material

775_2011_754_MOESM1_ESM.pdf (2.3 mb)
Supplementary material 1 (PDF 2403 kb)

References

  1. 1.
    Fermi G, Perutz MF, Shaanan B, Fourme R (1984) J Mol Biol 175:159–174PubMedCrossRefGoogle Scholar
  2. 2.
    Das TK, Couture M, Lee HC, Peisach J, Rousseau DL, Wittenberg BA, Wittenberg JB, Guertin M (1999) Biochemistry 38:15360–15368PubMedCrossRefGoogle Scholar
  3. 3.
    Fago A, Hundahl C, Malte H, Weber RE (2004) IUBMB Life 56:689–696PubMedCrossRefGoogle Scholar
  4. 4.
    de Sanctis D, Dewilde S, Vonrhein C, Pesce A, Moens L, Ascenzi P, Hankeln T, Burmester T, Ponassi M, Nardini M, Bolognesi M (2005) J Biol Chem 280:27222–27229PubMedCrossRefGoogle Scholar
  5. 5.
    Hoy JA, Hargrove MS (2008) Plant Physiol Biochem 46:371–379PubMedCrossRefGoogle Scholar
  6. 6.
    Pesce A, Thijs L, Nardini M, Desmet F, Sisinni L, Gourlay L, Bolli A, Coletta M, Van Doorslaer S, Wan X, Alam M, Ascenzi P, Moens L, Bolognesi M, Dewilde S (2009) J Mol Biol 386:246–260PubMedCrossRefGoogle Scholar
  7. 7.
    Vinogradov SN, Moens L (2008) J Biol Chem 283:8773–8777PubMedCrossRefGoogle Scholar
  8. 8.
    Scott NL, Xu Y, Shen G, Vuletich DA, Falzone CJ, Li Z, Ludwig M, Pond MP, Preimesberger MR, Bryant DA, Lecomte JTJ (2010) Biochemistry 49:7000–7011PubMedCrossRefGoogle Scholar
  9. 9.
    Smagghe BJ, Trent JT 3rd, Hargrove MS (2008) PLoS ONE 3:e2039PubMedCrossRefGoogle Scholar
  10. 10.
    Couture M, Das TK, Savard PY, Ouellet Y, Wittenberg JB, Wittenberg BA, Rousseau DL, Guertin M (2000) Eur J Biochem 267:4770–4780PubMedCrossRefGoogle Scholar
  11. 11.
    Scott NL, Falzone CJ, Vuletich DA, Zhao J, Bryant DA, Lecomte JTJ (2002) Biochemistry 41:6902–6910PubMedCrossRefGoogle Scholar
  12. 12.
    Pesce A, Couture M, Dewilde S, Guertin M, Yamauchi K, Ascenzi P, Moens L, Bolognesi M (2000) EMBO J 19:2424–2434PubMedCrossRefGoogle Scholar
  13. 13.
    Vu BC, Jones AD, Lecomte JTJ (2002) J Am Chem Soc 124:8544–8545PubMedCrossRefGoogle Scholar
  14. 14.
    Vu BC, Vuletich DA, Kuriakose SA, Falzone CJ, Lecomte JTJ (2004) J Biol Inorg Chem 9:183–194PubMedCrossRefGoogle Scholar
  15. 15.
    Goddard AD, Stevens JM, Rao F, Mavridou DA, Chan W, Richardson DJ, Allen JW, Ferguson SJ (2010) J Biol Chem 285:22882–22889PubMedCrossRefGoogle Scholar
  16. 16.
    Lee D, Pervushin K, Bischof D, Braun M, Thöny-Meyer L (2005) J Am Chem Soc 127:3716–3717PubMedCrossRefGoogle Scholar
  17. 17.
    Jiang Y, Trnka MJ, Medzihradszky KF, Ouellet H, Wang Y, Ortiz de Montellano PR (2009) J Inorg Biochem 103:316–325PubMedCrossRefGoogle Scholar
  18. 18.
    Metcalfe CL, Ott M, Patel N, Singh K, Mistry SC, Goff HM, Raven EL (2004) J Am Chem Soc 126:16242–16248PubMedCrossRefGoogle Scholar
  19. 19.
    Metcalfe CL, Daltrop O, Ferguson SJ, Raven EL (2007) Biochem J 408:355–361PubMedCrossRefGoogle Scholar
  20. 20.
    Scott NL, Lecomte JTJ (2000) Protein Sci 9:587–597PubMedCrossRefGoogle Scholar
  21. 21.
    de Duve C (1948) Acta Chem Scand 2:264–289CrossRefGoogle Scholar
  22. 22.
    Antonini E, Brunori M (1971) Hemoglobin and myoglobin in their reactions with ligands. North-Holland, AmsterdamGoogle Scholar
  23. 23.
    Teale FWJ (1959) Biochim Biophys Acta 35:543PubMedCrossRefGoogle Scholar
  24. 24.
    Englander SW, Calhoun DB, Englander JJ (1987) Anal Biochem 161:300–306PubMedCrossRefGoogle Scholar
  25. 25.
    Aitken CE, Marshall RA, Puglisi JD (2008) Biophys J 94:1826–1835PubMedCrossRefGoogle Scholar
  26. 26.
    Sun Y, Oberley LW (1989) Free Radic Biol Med 7:595–602PubMedCrossRefGoogle Scholar
  27. 27.
    Nelson DP, Kiesow LA (1972) Anal Biochem 49:474–478PubMedCrossRefGoogle Scholar
  28. 28.
    Di Iorio EE (1981) Methods Enzymol 76:57–72PubMedCrossRefGoogle Scholar
  29. 29.
    Henry ER, Hofrichter J (1992) Methods Enzymol 210:129–192CrossRefGoogle Scholar
  30. 30.
    Bilsel O, Zitzewitz JA, Bowers KE, Matthews CR (1999) Biochemistry 38:1018–1029PubMedCrossRefGoogle Scholar
  31. 31.
    Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) J Biomol NMR 6:277–293PubMedCrossRefGoogle Scholar
  32. 32.
    Goddard TD, Kneller DG (2006) Sparky. University of California, San FranciscoGoogle Scholar
  33. 33.
    Nothnagel HJ, Love N, Lecomte JT (2009) J Inorg Biochem 103:107–116PubMedCrossRefGoogle Scholar
  34. 34.
    La Mar GN, Satterlee JD, de Ropp JS (2000) In: Smith KM, Kadish K, Guilard R (eds) The porphyrin handbook. Academic Press, Burlington, pp 185–298Google Scholar
  35. 35.
    Hvitved AN, Trent JT 3rd, Premer SA, Hargrove MS (2001) J Biol Chem 276:34714–34721PubMedCrossRefGoogle Scholar
  36. 36.
    Bertini I, Luchinat C, Parigi G, Walker FA (1999) J Biol Inorg Chem 4:515–519PubMedCrossRefGoogle Scholar
  37. 37.
    Samuni U, Ouellet Y, Guertin M, Friedman JM, Yeh SR (2004) J Am Chem Soc 126:2682–2683PubMedCrossRefGoogle Scholar
  38. 38.
    Nonaka D, Wariishi H, Fujii H (2009) Biochemistry 48:898–905PubMedCrossRefGoogle Scholar
  39. 39.
    Egawa T, Shimada H, Ishimura Y (2000) J Biol Chem 275:34858–34866PubMedCrossRefGoogle Scholar
  40. 40.
    Cooper CE, Jurd M, Nicholls P, Wankasi MM, Svistunenko DA, Reeder BJ, Wilson MT (2005) Dalton Trans 3483–3488Google Scholar
  41. 41.
    Lardinois OM, de Montellano PR (2001) J Biol Chem 276:23186–23191PubMedCrossRefGoogle Scholar
  42. 42.
    Malencik DA, Anderson SR (2003) Amino Acids 25:233–247PubMedCrossRefGoogle Scholar
  43. 43.
    Huang L, Wojciechowski G, Ortiz de Montellano PR (2006) J Biol Chem 281:18983–18988PubMedCrossRefGoogle Scholar
  44. 44.
    Usha Devi S, Ramasarma T (1987) Mol Cell Biochem 77:111–120PubMedGoogle Scholar
  45. 45.
    Netto LE, Stadtman ER (1996) Arch Biochem Biophys 333:233–242PubMedCrossRefGoogle Scholar
  46. 46.
    Hoy JA, Smagghe BJ, Halder P, Hargrove MS (2007) Protein Sci 16:250–260PubMedCrossRefGoogle Scholar
  47. 47.
    Paul KG, Theorell H, Akeson A (1953) Acta Chem Scan 7:1284–1287CrossRefGoogle Scholar
  48. 48.
    Barker PD, Ferrer JC, Mylrajan M, Loehr TM, Feng R, Konishi Y, Funk WD, MacGillivray RT, Mauk AG (1993) Proc Natl Acad Sci USA 90:6542–6546PubMedCrossRefGoogle Scholar
  49. 49.
    Falzone CJ, Vu BC, Scott NL, Lecomte JTJ (2002) J Mol Biol 324:1015–1029PubMedCrossRefGoogle Scholar
  50. 50.
    Marzocchi MP, Smulevich G (2003) J Raman Spectrosc 34:725–736CrossRefGoogle Scholar
  51. 51.
    Satterlee JD, Erman JE (1983) J Biol Chem 258:1050–1056PubMedGoogle Scholar
  52. 52.
    Barrows TP, Poulos TL (2005) Biochemistry 44:14062–14068PubMedCrossRefGoogle Scholar
  53. 53.
    Guallar V, Olsen B (2006) J Inorg Biochem 100:755–760PubMedCrossRefGoogle Scholar
  54. 54.
    Xu C, Ibrahim M, Spiro TG (2008) Biochemistry 47:2379–2387PubMedCrossRefGoogle Scholar
  55. 55.
    Hoy JA, Kundu S, Trent JT 3rd, Ramaswamy S, Hargrove MS (2004) J Biol Chem 279:16535–16542PubMedCrossRefGoogle Scholar
  56. 56.
    Trent JT 3rd, Kundu S, Hoy JA, Hargrove MS (2004) J Mol Biol 341:1097–1108PubMedCrossRefGoogle Scholar
  57. 57.
    Trevaskis B, Watts RA, Andersson CR, Llewellyn DJ, Hargrove MS, Olson JS, Dennis ES, Peacock WJ (1997) Proc Natl Acad Sci USA 94:12230–12234PubMedCrossRefGoogle Scholar
  58. 58.
    Burmester T, Ebner B, Weich B, Hankeln T (2002) Mol Biol Evol 19:416–421PubMedGoogle Scholar
  59. 59.
    Sturms R, Kakar S, Trent J 3rd, Hargrove MS (2010) Biochemistry 49:4085–4093PubMedCrossRefGoogle Scholar
  60. 60.
    Hargrove MS (2000) Biophys J 79:2733–2738PubMedCrossRefGoogle Scholar
  61. 61.
    Weiland TR, Kundu S, Trent JT 3rd, Hoy JA, Hargrove MS (2004) J Am Chem Soc 126:11930–11935PubMedCrossRefGoogle Scholar
  62. 62.
    Yoon J, Herzik MA, Winter MB, Tran R, Olea C, Marletta MA (2010) Biochemistry 49:5662–5670Google Scholar
  63. 63.
    Lardinois OM, Tomer KB, Mason RP, Deterding LJ (2008) Biochemistry 47:10440–10448PubMedCrossRefGoogle Scholar
  64. 64.
    Hamdane D, Vasseur-Godbillon C, Baudin-Creuza V, Hoa GH, Marden MC (2007) J Biol Chem 282:6398–6404PubMedCrossRefGoogle Scholar
  65. 65.
    Goodrich LE, Paulat F, Praneeth VK, Lehnert N (2010) Inorg Chem 49:6293–6316PubMedCrossRefGoogle Scholar
  66. 66.
    Minamizaki K, Mizoguchi T, Goto T, Tamiaki H, Fujita Y (2008) J Biol Chem 283:2684–2692PubMedCrossRefGoogle Scholar
  67. 67.
    Yilmaz M, Kang I, Beale SI (2010) Photosynth Res 103:47–59PubMedCrossRefGoogle Scholar
  68. 68.
    Cowley AB, Kennedy ML, Silchenko S, Lukat-Rodgers GS, Rodgers KR, Benson DR (2006) Inorg Chem 45:9985–10001PubMedCrossRefGoogle Scholar
  69. 69.
    Smagghe BJ, Halder P, Hargrove MS (2008) Methods Enzymol 436:359–378PubMedCrossRefGoogle Scholar
  70. 70.
    Smagghe BJ, Sarath G, Ross E, Hilbert JL, Hargrove MS (2006) Biochemistry 45:561–570PubMedCrossRefGoogle Scholar
  71. 71.
    Stevens JM, Daltrop O, Higham CW, Ferguson SJ (2003) J Biol Chem 278:20500–20506PubMedCrossRefGoogle Scholar
  72. 72.
    Daltrop O, Allen JW, Willis AC, Ferguson SJ (2002) Proc Natl Acad Sci USA 99:7872–7876PubMedCrossRefGoogle Scholar
  73. 73.
    Kranz RG, Richard-Fogal C, Taylor JS, Frawley ER (2009) Microbiol Mol Biol Rev 73:510–528PubMedCrossRefGoogle Scholar
  74. 74.
    Schneider S, Marles-Wright J, Sharp KH, Paoli M (2007) Nat Prod Rep 24:621–630PubMedCrossRefGoogle Scholar
  75. 75.
    Shelnutt JA, Song XZ, Ma JG, Jia SL, Jentzen W, Medforth CJ (1998) Chem Soc Rev 27:31–41CrossRefGoogle Scholar
  76. 76.
    Cowley AB, Lukat-Rodgers GS, Rodgers KR, Benson DR (2004) Biochemistry 43:1656–1666PubMedCrossRefGoogle Scholar
  77. 77.
    Bowman SE, Bren KL (2010) Inorg Chem 49:7890–7897PubMedCrossRefGoogle Scholar
  78. 78.
    Allen JW, Ginger ML, Ferguson SJ (2004) Biochem J 383:537–542PubMedCrossRefGoogle Scholar
  79. 79.
    Kurisu G, Zhang H, Smith JL, Cramer WA (2003) Science 302:1009–1014PubMedCrossRefGoogle Scholar

Copyright information

© SBIC 2011

Authors and Affiliations

  • Henry J. Nothnagel
    • 1
  • Matthew R. Preimesberger
    • 1
  • Matthew P. Pond
    • 1
  • Benjamin Y. Winer
    • 1
  • Emily M. Adney
    • 1
  • Juliette T. J. Lecomte
    • 1
  1. 1.T.C. Jenkins Department of BiophysicsJohns Hopkins UniversityBaltimoreUSA

Personalised recommendations