Spectroscopic analyses of 2-oxoglutarate-dependent oxygenases: TauD as a case study

  • Denis A. Proshlyakov
  • John McCracken
  • Robert P. Hausinger
Part of the following topical collections:
  1. 60 Years of Oxygen Activation


A wide range of spectroscopic approaches have been used to interrogate the mononuclear iron metallocenter in 2-oxoglutarate (2OG)-dependent oxygenases. The results from these spectroscopic studies have provided valuable insights into the structural changes at the active site during substrate binding and catalysis, thus providing critical information that complements investigations of these enzymes by X-ray crystallography, biochemical, and computational approaches. This mini-review highlights taurine hydroxylase (taurine:2OG dioxygenase, TauD) as a case study to illustrate the wealth of knowledge that can be generated by applying a diverse array of spectroscopic investigations to a single enzyme. In particular, electronic absorption, circular dichroism, magnetic circular dichroism, conventional and pulse electron paramagnetic, Mössbauer, X-ray absorption, and resonance Raman methods have been exploited to uncover the properties of the metal site in TauD.


2-Oxoglutarate Nonheme iron Oxygenase Spectroscopy 



Circular dichroism


Continuous flow


Continuous wave




Electron paramagnetic resonance


Electron spin echo envelope modulation


Extended X-ray absorption fine structure


Freeze quench


Fourier transform


Hyperfine sublevel correlation


Ligand-to-metal charge-transfer


Magnetic circular dichroism


Metal-to-ligand charge-transfer




Resonance Raman




Taurine:2OG dioxygenase




X-ray absorption spectroscopy



Work in the authors’ laboratories on this topic was supported by the National Institutes of Health (GM096132 to DAP, GM054065 and RR15880 to JM, and GM063584 to RPH).


  1. 1.
    Hausinger RP (2015) In: Schofield CJ, Hausinger RP (eds) 2-Oxoglutarate-dependent oxygenases. Royal Society of Chemistry, Cambridge, pp 1–58Google Scholar
  2. 2.
    Wu L-F, Meng S, Tang G-L (2016) Biochim Biophys Acta 1864:453–470CrossRefPubMedGoogle Scholar
  3. 3.
    Martinez S, Hausinger RP (2015) J Biol Chem 290:20702–20711CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Aik WS, Chowdhury R, Clifton IJ, Hopkinson RJ, Leissing T, McDonough MA, Nowak R, Schofield CJ, Walport LJ (2015) In: Schofield CJ, Hausinger RP (eds) 2-oxoglutarate-dependent oxygenases. Royal Society of Chemistry, CambridgeGoogle Scholar
  5. 5.
    Proshlyakov DA, Hausinger RP (2011) In: Kumar D, de Visser SP (eds) Iron-containing enzymes: versatile catalysts of hydroxylation reactions in nature. Royal Society of Chemistry, Cambridge, pp 67–87CrossRefGoogle Scholar
  6. 6.
    Eichhorn E, van der Ploeg JR, Kertesz MA, Leisinger T (1997) J Biol Chem 272:23031–23036CrossRefPubMedGoogle Scholar
  7. 7.
    Grzyska PK, Hausinger RP, Proshlyakov DA (2010) Anal Biochem 399:64–71CrossRefPubMedGoogle Scholar
  8. 8.
    Ryle MJ, Padmakumar R, Hausinger RP (1999) Biochemistry 38:15278–15286CrossRefPubMedGoogle Scholar
  9. 9.
    Ryle MJ, Liu A, Muthukumaran RB, Ho RYN, Koehntop KD, McCracken J, Que L Jr, Hausinger RP (2003) Biochemistry 42:1854–1862CrossRefPubMedGoogle Scholar
  10. 10.
    Price JC, Barr EW, Tirupati B, Bollinger JM Jr, Krebs C (2003) Biochemistry 42:7497–7508CrossRefPubMedGoogle Scholar
  11. 11.
    Grzyska PK, Ryle MJ, Monterosso GR, Liu J, Ballou DP, Hausinger RP (2005) Biochemistry 44:3845–3855CrossRefPubMedGoogle Scholar
  12. 12.
    Knauer SH, Hartl-Spiegelhauer O, Schwarzinger S, Hänzelmann P, Dobbek H (2012) FEBS J 279:816–831CrossRefPubMedGoogle Scholar
  13. 13.
    Pavel EG, Zhou J, Busby RW, Gunsior M, Townsend CA, Solomon EI (1998) J Am Chem Soc 120:743–753CrossRefGoogle Scholar
  14. 14.
    Simmons JM, Koslowski DJ, Hausinger RP (2010) Exp Parasitol 124:453–458CrossRefPubMedGoogle Scholar
  15. 15.
    Ryle MJ, Koehntop KD, Liu A, Que L Jr, Hausinger RP (2003) Proc Natl Acad Sci USA 100:3790–3795CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Koehntop KD, Marimanikkuppam S, Ryle MJ, Hausinger RP, Que L Jr (2006) J Biol Inorg Chem 11:63–72CrossRefPubMedGoogle Scholar
  17. 17.
    Grzyska PK, Müller TA, Campbell MG, Hausinger RP (2007) J Inorg Biochem 101:797–808CrossRefPubMedGoogle Scholar
  18. 18.
    Grzyska PK, Hausinger RP (2007) Inorg Chem 46:10087–10092CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Liu A, Ho RYN, Que L Jr, Ryle MJ, Phinney BS, Hausinger RP (2001) J Am Chem Soc 123:5126–5127CrossRefPubMedGoogle Scholar
  20. 20.
    Henshaw TF, Feig M, Hausinger RP (2004) J Inorg Biochem 98:856–861CrossRefPubMedGoogle Scholar
  21. 21.
    Chen Y-H, Comeaux LM, Eyles SJ, Knapp MJ (2008) Chem Commun 4768–4770Google Scholar
  22. 22.
    Yan W, Song H, Song F, Guo Y, Wu CH, Her AS, Pu Y, Wang S, Naowarojna N, Weitz A, Hendrich MP, Costello CE, Zhang L, Liu P, Zhang YJ (2015) Nature 527:539–543CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Price JC, Barr EW, Hoffart LM, Krebs C, Bollinger JM Jr (2005) Biochemistry 44:8138–8147CrossRefPubMedGoogle Scholar
  24. 24.
    Price JC, Barr EW, Glass TE, Krebs C, Bollinger JM Jr (2003) J Am Chem Soc 125:13008–13009CrossRefPubMedGoogle Scholar
  25. 25.
    McCusker KP, Klinman JP (2009) Proc Natl Acad Sci USA 106:19791–19795CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    McCusker KP, Klinman JP (2010) J Am Chem Soc 132:5114–5120CrossRefPubMedGoogle Scholar
  27. 27.
    Hoffart LM, Barr EW, Guyer RB, Bollinger JM Jr, Krebs C (2006) Proc Natl Acad Sci USA 103:14738–14743CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Matthews MM, Krest C, Barr EW, Vaillancourt F, Walsh CT, Green M, Krebs C, Bollinger JM Jr (2009) Biochemistry 48:4331–4343CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Galonić DP, Barr EW, Walsh CT, Bollinger JM Jr, Krebs C (2007) Nat Chem Biol 3:113–116CrossRefPubMedGoogle Scholar
  30. 30.
    Solomon EI (2001) Inorg Chem 40:3656–3669CrossRefPubMedGoogle Scholar
  31. 31.
    Solomon EI, Light KM, Liu LV, Srnec M, Wong SD (2013) Acc Chem Res 46:2725–2739CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Solomon EI, Brunold TC, Davis MI, Kemsley JN, Lee S-K, Lehnert N, Neese F, Skulan AJ, Yang Y-S, Zhou J (2000) Chem Rev 100:235–349CrossRefPubMedGoogle Scholar
  33. 33.
    Neidig ML, Brown CD, Light KM, Fujimori DG, Nolan EM, Price JC, Barr EW, Bollinger JM Jr, Krebs C, Walsh CT, Solomon EI (2007) J Am Chem Soc 129:14224–14231CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Zhou J, Kelly WL, Bachmann BO, Gunsior M, Townsend CA, Solomon EI (2001) J Am Chem Soc 123:7388–7398CrossRefPubMedGoogle Scholar
  35. 35.
    Kemsley JN, Mitic N, Zaleski L, Caradonna JP, Solomon EI (1999) J Am Chem Soc 121:1528–1536CrossRefGoogle Scholar
  36. 36.
    Chow MS, Eser BE, Wilson SA, Hodgson KO, Hedman B, Fitzpatrick PF, Solomon EI (2009) J Am Chem Soc 131:7685–7698CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Enemark JH, Feltham RD (1974) Coord Chem Rev 13:339–406CrossRefGoogle Scholar
  38. 38.
    Arciero DM, Lipscomb JD, Huynh BH, Kent TA, Münck E (1983) J Biol Chem 258:14981–14991PubMedGoogle Scholar
  39. 39.
    Orville AM, Chen VJ, Kriauciunas A, Harpel MR, Fox BG, Münck E, Lipscomb JD (1992) Biochemistry 31:4602–4612CrossRefPubMedGoogle Scholar
  40. 40.
    Chen VJ, Orville AM, Harpel MR, Frolik CA, Surerus KK, Münck E, Lipscomb JD (1989) J Biol Chem 264:21677–21681PubMedGoogle Scholar
  41. 41.
    Rocklin AM, Tierney DL, Kofman V, Brunhuber NMW, Hoffman BM, Christoffersen RE, Reich NO, Lipscomb JD, Que L Jr (1999) Proc Natl Acad Sci USA 96:7905–7909CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Tierney DL, Rocklin AM, Lipscomb JD, Que L Jr, Hoffman B (2005) J Am Chem Soc 127:7005–7013CrossRefPubMedGoogle Scholar
  43. 43.
    Yang TC, Wolfe MD, Neibergall MB, Mekmouche Y, Lipscomb JD, Hoffman BM (2003) J Am Chem Soc 125:7056–7066CrossRefPubMedGoogle Scholar
  44. 44.
    Brown CA, Pavlovsky MA, Westre TE, Zhang Y, Hedman B, Hodgson KO, Solomon EI (1995) J Am Chem Soc 117:715–732CrossRefGoogle Scholar
  45. 45.
    Aquino F, Rodriguez JH (2009) J Phys Chem A 113:9150–9156CrossRefPubMedGoogle Scholar
  46. 46.
    Muthukumaran RB, Grzyska PK, Hausinger RP, McCracken J (2007) Biochemistry 46:5951–5959CrossRefPubMedGoogle Scholar
  47. 47.
    McCracken J (2015) Meth Enzymol 563:285–309CrossRefPubMedGoogle Scholar
  48. 48.
    Casey TM, Grzyska PK, Hausinger RP, McCracken J (2013) J Phys Chem B 117:10384–10394CrossRefPubMedGoogle Scholar
  49. 49.
    Stoll S, Britt RD (2009) Phys Chem Chem Phys 11:6614–6625CrossRefPubMedGoogle Scholar
  50. 50.
    Stoll S, Schweiger A (2006) J Magn Res 178:42–55CrossRefGoogle Scholar
  51. 51.
    Krzyaniak MD, Eser BE, Ellis HR, Fitzpatrick PF, McCracken J (2013) Biochemistry 52:8430–8441CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Elkins JM, Ryle MJ, Clifton IJ, Dunning Hotopp JC, Lloyd JS, Burzlaff NI, Baldwin JE, Hausinger RP, Roach PL (2002) Biochemistry 41:5185–5192CrossRefPubMedGoogle Scholar
  53. 53.
    Shane JJ, Höfer P, Reijerse EJ, de Boer E (1992) J Magn Res 99:596–604Google Scholar
  54. 54.
    McCracken J, Cappillino PJ, McNally JS, Krzyaniak MD, Howart M, Tarves PC, Caradonna JP (2015) Inorg Chem 54:6486–6497CrossRefPubMedGoogle Scholar
  55. 55.
    McCracken J, Eser BE, Mannikko D, Krzyaniak MD, Fitzpatrick PF (2015) Biochemistry 54:3759–3771CrossRefPubMedGoogle Scholar
  56. 56.
    Krebs C, Price JC, Baldwin J, Saleh L, Green MT, Bollinger JM Jr (2005) Inorg Chem 44:742–757CrossRefPubMedGoogle Scholar
  57. 57.
    Sinnecker S, Svensen N, Barr EW, Ye S, Bollinger JM Jr, Neese F, Krebs C (2007) J Am Chem Soc 129:6168–6179CrossRefPubMedGoogle Scholar
  58. 58.
    Chang W-C, Guo Y, Wang C, Butch SE, Rosenzweig AC, Boal AK, Krebs C, Bollinger JM Jr (2014) Science 343:1140–1144CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Chang W-C, Li J, Lee JL, Cronican AA, Guo Y (2016) J Am Chem Soc 138:10390–10393CrossRefPubMedGoogle Scholar
  60. 60.
    Riggs-Gelasco PJ, Price JC, Guyer RB, Brehm JH, Barr EW, Bollinger JM Jr, Krebs C (2004) J Am Chem Soc 126:8108–8109CrossRefPubMedGoogle Scholar
  61. 61.
    Cosper NJ, Stälhandske CMV, Saari RE, Hausinger RP, Scott RA (1999) J Biol Inorg Chem 4:122–129CrossRefPubMedGoogle Scholar
  62. 62.
    Giri NC, Sung H, Chen H, Costa M, Maroney MJ (2011) Biochemistry 50:5067–5076CrossRefPubMedPubMedCentralGoogle Scholar
  63. 63.
    Lloyd MD, Lee H-J, Harlos K, Zhang Z-H, Baldwin JE, Schofield CJ, Charnock JM, Garner CD, Hara T, TerwisschaVanScheltinga AC, Valegård K, Viklund JAC, Hajdu J, Andersson I, Danielsson A, Bhikhabhai R (1999) J Mol Biol 287:943–960CrossRefPubMedGoogle Scholar
  64. 64.
    Fujimori DG, Barr EW, Matthews ML, Koch GM, Yonce JR, Walsh CT, Bollinger JM Jr, Krebs C, Riggs-Gelasco PJ (2007) J Am Chem Soc 129:13408–13409CrossRefPubMedGoogle Scholar
  65. 65.
    Ye S, Price JC, Barr EW, Green MT, Bollinger JM Jr, Krebs C, Neese F (2010) J Am Chem Soc 132:4739–4751CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Ho RYN, Mehn MP, Hegg EL, Liu A, Ryle MA, Hausinger RP, Que L Jr (2001) J Am Chem Soc 123:5022–5029CrossRefPubMedGoogle Scholar
  67. 67.
    Proshlyakov DA, Henshaw TF, Monterosso GR, Ryle MJ, Hausinger RP (2004) J Am Chem Soc 126:1022–1023CrossRefPubMedGoogle Scholar
  68. 68.
    Grzyska PK, Appelman EH, Hausinger RP, Proshlyakov DA (2010) Proc Natl Acad Sci USA 107:3982–3987CrossRefPubMedPubMedCentralGoogle Scholar
  69. 69.
    Bell RP (1980) The tunnel effect in chemistry. Chapman & Hall, LondonCrossRefGoogle Scholar
  70. 70.
    Krishtalik LI (1979) Charge transfer in reactions in electrochemical and chemical processes. Plenum, New YorkGoogle Scholar
  71. 71.
    Kuznetsov AM, Ulstrup J (2006) In: Cohen A, Limbach H-H (eds) Isotope effects in chemistry and biology. CRC Press, Boca Raton, pp 691–724Google Scholar
  72. 72.
    Mayer JM (2011) Acc Chem Res 44:36–46CrossRefPubMedGoogle Scholar
  73. 73.
    Brischwein M, Scharf B, Engelhard M, Mäntele W (1993) Biochemistry 32:13710–13717CrossRefPubMedGoogle Scholar
  74. 74.
    Hirst J (2006) Biochim Biophys Acta 1757:225–239CrossRefPubMedGoogle Scholar
  75. 75.
    Costentin C, Robert M, Saveant JM (2010) Chem Rev 110:PR1–PR40Google Scholar
  76. 76.
    Sitter AJ, Reczek CM, Terner J (1985) J Biol Chem 260:7515–7522PubMedGoogle Scholar
  77. 77.
    Hashimoto S, Teraoka J, Inubushi T, Yonetani T, Kitagawa T (1986) J Biol Chem 261:11110–11118PubMedGoogle Scholar
  78. 78.
    Proshlyakov DA, Ogura T, Shinzawa-Itoh K, Yoshikawa S, Appelman EH, Kitagawa T (1994) J Biol Chem 269:29385–29388PubMedGoogle Scholar
  79. 79.
    Ogo S, Yamahara R, Roach M, Suenobu T, Aki M, Ogura T, Kitagawa T, Masuda H, Fukuzumi S, Watanabe Y (2002) Inorg Chem 41:5513–5520CrossRefPubMedGoogle Scholar
  80. 80.
    Wang H, Alp EE, Yoda Y, Cramer SP (2014) Methods Mol Biol 1122:125–137CrossRefPubMedPubMedCentralGoogle Scholar
  81. 81.
    Sage JT, Paxson C, Wyllie GRA, Sturhahn W, Durbin SM, Champion PM, Alp EE, Scheidt WR (2001) J Phys Condens Matter 13:7707–7722CrossRefGoogle Scholar
  82. 82.
    Wong SD, Srnec M, Matthews ML, Liu LV, Kwak Y, Park K, Bell CB III, Alp EE, Zhao J, Yoda Y, Kitao S, Seto M, Krebs C, Bollinger JM Jr, Solomon EI (2013) Nature 499:320–323CrossRefPubMedPubMedCentralGoogle Scholar
  83. 83.
    O’Brien JR, Schuller DJ, Yang VS, Dillard BD, Lanzilotta WN (2003) Biochemistry 42:5547–5554CrossRefPubMedGoogle Scholar
  84. 84.
    Henderson KL, Müller TA, Hausinger RP, Emerson JP (2015) Inorg Chem 54:2278–2283CrossRefPubMedGoogle Scholar
  85. 85.
    Mirica LM, McCusker KP, Munos JW, Liu H-W, Klinman JP (2008) J Am Chem Soc 130:8122–8123CrossRefPubMedPubMedCentralGoogle Scholar
  86. 86.
    de Visser SP (2006) J Am Chem Soc 128:9813–9824CrossRefPubMedGoogle Scholar
  87. 87.
    de Visser SP (2007) Chem Commun 171–173Google Scholar
  88. 88.
    Godfrey E, Porro CS, de Visser SP (2008) J Phys Chem A 112:2464–2468CrossRefPubMedGoogle Scholar
  89. 89.
    De Visser SP, Quesne MG, Martin CB, Comba P, Ryde U (2014) Chem Commun 50:262–282CrossRefGoogle Scholar
  90. 90.
    Borowski T, Bassan A, Siegbahn PEM (2004) Chem Eur J 10:1031–1041CrossRefPubMedGoogle Scholar
  91. 91.
    Shaik S, Hirao H, Kumar D (2007) Acc Chem Res 40:532–542CrossRefPubMedGoogle Scholar
  92. 92.
    Cho K-B, Hirao H, Shaik S, Nam W (2016) Chem Soc Rev 45:1197–1210CrossRefPubMedGoogle Scholar
  93. 93.
    Geng C, Ye S, Neese F (2014) Dalton Trans 43:6079–6086CrossRefPubMedGoogle Scholar
  94. 94.
    Allpress CJ, Kleespies ST, Que L Jr (2015) In: Schofield CJ, Hausinger RP (eds) 2-oxoglutarate-dependent oxygenases. Royal Society of Chemistry, Cambridge, pp 123–148Google Scholar
  95. 95.
    Puri M, Que L Jr (2015) Acc Chem Res 48:2443–2452CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© SBIC 2016

Authors and Affiliations

  • Denis A. Proshlyakov
    • 1
  • John McCracken
    • 1
  • Robert P. Hausinger
    • 2
    • 3
  1. 1.Department of ChemistryMichigan State UniversityEast LansingUSA
  2. 2.Department of Microbiology and Molecular GeneticsMichigan State UniversityEast LansingUSA
  3. 3.Department of Biochemistry and Molecular BiologyMichigan State UniversityEast LansingUSA

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