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JBIC Journal of Biological Inorganic Chemistry

, Volume 23, Issue 7, pp 1093–1104 | Cite as

Mn(III) species formed by the multi-copper oxidase MnxG investigated by electron paramagnetic resonance spectroscopy

  • Lizhi Tao
  • Troy A. Stich
  • Alexandra V. Soldatova
  • Bradley M. Tebo
  • Thomas G. Spiro
  • William H. Casey
  • R. David BrittEmail author
Original Paper
Part of the following topical collections:
  1. Alison Butler: Papers in Celebration of Her 2018 ACS Alfred Bader Award in Bioorganic or Bioinorganic Chemistry

Abstract

The multi-copper oxidase (MCO) MnxG from marine Bacillus bacteria plays an essential role in geochemical cycling of manganese by oxidizing Mn2+(aq) to form manganese oxide minerals at rates that are three to five orders of magnitude faster than abiotic rates. The MCO MnxG protein is isolated as part of a multi-protein complex, denoted as Mnx, which includes one MnxG unit and a hexamer of MnxE3F3 subunit. During the oxidation of Mn2+(aq) catalyzed by the Mnx protein complex, an enzyme-bound Mn(III) species was trapped recently in the presence of pyrophosphate (PP) and analyzed using parallel-mode electron paramagnetic resonance (EPR) spectroscopy. Herein, we provide a full analysis of this enzyme-bound Mn(III) intermediate via temperature dependence studies and spectral simulations. This Mnx-bound Mn(III) species is characterized by a hyperfine-coupling value of A(55Mn) = 4.2 mT (corresponding to 120 MHz) and a negative zero-field splitting (ZFS) value of D = − 2.0 cm−1. These magnetic properties suggest that the Mnx-bound Mn(III) species could be either six-coordinate with a 5B1g ground state or square-pyramidal five-coordinate with a 5B1 ground state. In addition, as a control, Mn(III)PP is also analyzed by parallel-mode EPR spectroscopy. It exhibits distinctly different magnetic properties with a hyperfine-coupling value of A(55Mn) = 4.8 mT (corresponding to 140 MHz) and a negative ZFS value of D = − 2.5 cm−1. The different ZFS values suggest differences in ligand environment of Mnx-bound Mn(III) and aqueous Mn(III)PP species. These studies provide further insights into the mechanism of biological Mn2+(aq) oxidation.

Keywords

Parallel-mode EPR Multi-copper oxidase MnxG Mnx protein complex Mn(II) oxidation Zero-field splitting 

Notes

Acknowledgements

The work was supported by the National Science Foundation Award Numbers CHE-1213699, CHE-1665455 to RDB, EAR-1231322 to WHC, CHE-1410688 to BMT, and CHE-1410353 to TGS. The EPR spectrometers at the CalEPR facility used in this study were funded by the National Institutes of Health (S10-RR021075) and the NSF (CHE-1048671).

References

  1. 1.
    Post JE (1999) Manganese oxide minerals: crystal structures and economic and environmental significance. Proc Natl Acad Sci USA 96(7):3447–3454CrossRefGoogle Scholar
  2. 2.
    Tebo BM, Bargar JR, Clement BG, Dick GJ, Murray KJ, Parker D, Verity R, Webb SM (2004) Biogenic manganese oxides: properties and mechanisms of formation. Annu Rev Earth Planet Sci 32(1):287–328CrossRefGoogle Scholar
  3. 3.
    Spiro TG, Bargar JR, Sposito G, Tebo BM (2010) Bacteriogenic manganese oxides. Acc Chem Res 43(1):2–9CrossRefGoogle Scholar
  4. 4.
    Waite TD, Szymczak R (1994) Photoredox transformation of iron and manganese in marine systems: review of recent field investigations. Lewis Publishers, LondonGoogle Scholar
  5. 5.
    Sunda WG, Huntsman SA (1988) Effect of sunlight on redox cycles of manganese in the southwestern Sargasso Sea. Deep Sea Res 35(8):1297–1317CrossRefGoogle Scholar
  6. 6.
    Morgan JJ (2005) Kinetics of reaction between O2 and Mn(II) species in aqueous solutions. Geochim Cosmochim Acta 69(1):35–48CrossRefGoogle Scholar
  7. 7.
    Butterfield CN, Soldatova AV, Lee SW, Spiro TG, Tebo BM (2013) Mn(II, III) oxidation and MnO2 mineralization by an expressed bacterial multicopper oxidase. Proc Natl Acad Sci USA 110(29):11731–11735CrossRefGoogle Scholar
  8. 8.
    Waasbergen LG, Hildebrand M, Tebo BM (1996) Identification and characterization of a gene cluster involved in manganese oxidation by spores of the marine Bacillus sp. strain SG-1. J Bacteriol 178(12):3517–3530CrossRefGoogle Scholar
  9. 9.
    Dick GJ, Torpey JW, Beveridge TJ, Tebo BM (2008) Direct identification of a bacterial manganese(II) oxidase, the multicopper oxidase MnxG, from spores of several different marine Bacillus species. Appl Environ Microbiol 74(5):1527–1534CrossRefGoogle Scholar
  10. 10.
    Butterfield CN, Tao L, Chacon KN, Spiro TG, Blackburn NJ, Casey WH, Britt RD, Tebo BM (2015) Multicopper manganese oxidase accessory proteins bind Cu and Heme. Biochim Biophys Acta 1854(12):1853–1859CrossRefGoogle Scholar
  11. 11.
    Solomon EI, Szilagyi RK, George SD, Basumallick L (2004) Electronic structures of metal sites in proteins and models: contributions to function in blue copper proteins. Chem Rev 104(2):419–458CrossRefGoogle Scholar
  12. 12.
    Solomon EI, Sundaram UM, Machonkin T (1996) Multicopper oxidases and oxygenases. Chem Rev 96(7):2563–2605CrossRefGoogle Scholar
  13. 13.
    Lee SK, George SD, Antholine WE, Hedman B, Hodgson KO, Solomon EI (2002) Nature of the intermediate formed in the reduction of O2 to H2O at the Trinuclear Copper Cluster active site in native Laccase. J Am Chem Soc 124(21):6180–6193CrossRefGoogle Scholar
  14. 14.
    Solomon EI, Augustine AJ, Yoon J (2008) O2 reduction to H2O by the multicopper oxidases. Dalton Trans 30:3921–3932CrossRefGoogle Scholar
  15. 15.
    Heppner DE, Kjaergaard CH, Solomon EI (2013) Molecular origin of rapid versus slow intramolecular electron transfer in the catalytic cycle of the multicopper oxidases. J Am Chem Soc 135(33):12212–12215CrossRefGoogle Scholar
  16. 16.
    Quintanar L, Gebhard M, Wang TP, Kosman DJ, Solomon EI (2004) Ferrous binding to the multicopper oxidases Saccharomyces cerevisiae Fet3p and human ceruloplasmin: contributions to ferroxidase activity. J Am Chem Soc 126(21):6579–6589CrossRefGoogle Scholar
  17. 17.
    Machonkin TE, Solomon EI (2000) The thermodynamics, kinetics, and molecular mechanism of intramolecular electron transfer in human ceruloplasmin. J Am Chem Soc 122(50):12547–12560CrossRefGoogle Scholar
  18. 18.
    Lindley PF, Card G, Zaitseva I, Zaitsev V, Reinhammar B, Selin-Lindgren E, Yoshida K (1997) An X-ray structural study of human ceruloplasmin in relation to ferroxidase activity. J Biol Inorg Chem 2(4):454–463CrossRefGoogle Scholar
  19. 19.
    Singh SK, Roberts SA, McDevitt SF, Weichsel A, Wildner GF, Grass GB, Rensing C, Montfort WR (2011) Crystal structures of multicopper oxidase CueO bound to Copper(I) and Silver(I): functional role of a methionine-rich sequence. J Biol Chem 286(43):37849–37857CrossRefGoogle Scholar
  20. 20.
    Tao L, Stich TA, Butterfield CN, Romano C, Tebo BM, Casey WH, Britt RD (2015) Mn(II) binding and subsequent oxidation by the multicopper oxidase Mnx investigated by electron paramagnetic resonance spectroscopy. J Am Chem Soc 137(33):10563–10575CrossRefGoogle Scholar
  21. 21.
    Su J, Deng L, Huang L, Guo S, Liu F, He J (2014) Catalytic oxidation of manganese(II) by multicopper oxidase CueO and characterization of the biogenic Mn oxide. Water Res 56:304–313CrossRefGoogle Scholar
  22. 22.
    Su J, Bao P, Bai T, Deng L, Wu H, Liu F, He J (2013) CotA, a multicopper oxidase from Bacillus pumilus WH4, exhibits manganese-oxidase activity. PLoS One 8:e60573CrossRefGoogle Scholar
  23. 23.
    Romano CA, Zhou M, Song Y, Wysocki VH, Dohnalkova AC, Kovarik L, Paša-Tolić L, Tebo BM (2017) Biogenic manganese oxide nanoparticle formation by a multimeric multicopper oxidase Mnx. Nat Commun 8(1):746CrossRefGoogle Scholar
  24. 24.
    Tao L, Stich TA, Liou S-H, Soldatova AV, Delgadillo DA, Romano CA, Spiro TG, Goodin DB, Tebo BM, Casey WH, Britt RD (2017) Copper binding sites in the manganese-oxidizing Mnx protein complex investigated by electron paramagnetic resonance spectroscopy. J Am Chem Soc 139(26):8868–8877CrossRefGoogle Scholar
  25. 25.
    Tao L, Simonov AN, Romano CA, Butterfield CN, Fekete M, Tebo BM, Bond AM, Spiccia L, Martin LL, Casey WH (2017) Biogenic manganese-oxide mineralisation is enhanced by an oxidative priming mechanism for the multi-copper oxidase, MnxEFG. Chem Eur J 23(6):1346–1352CrossRefGoogle Scholar
  26. 26.
    Tao L, Simonov AN, Romano CA, Butterfield CN, Tebo BM, Bond AM, Spiccia L, Martin LL, Casey WH (2018) Probing electron transfer in the manganese-oxide-forming MnxEFG protein complex using Fourier transformed AC voltammetry: understanding the oxidative priming effect. ChemElectroChem 5(6):872–876CrossRefGoogle Scholar
  27. 27.
    Soldatova AV, Tao L, Romano CA, Stich TA, Casey WH, Britt RD, Tebo BM, Spiro TG (2017) Mn(II) oxidation by the multicopper oxidase complex Mnx: a binuclear activation mechanism. J Am Chem Soc 139(33):11369–11380CrossRefGoogle Scholar
  28. 28.
    Mathur P, Crowder M, Dismukes GC (1987) Dimanganese(II) complexes of a septadentate ligand. Functional analogs of the manganese pseudocatalase. J Am Chem Soc 109(17):5227–5233CrossRefGoogle Scholar
  29. 29.
    Soldatova AV, Romano CA, Tao L, Stich TA, Casey WH, Britt RD, Tebo BM, Spiro TG (2017) Mn(II) oxidation by the multicopper oxidase complex Mnx: a coordinated two-stage Mn(II)/(III) and Mn(III)/(IV) mechanism. J Am Chem Soc 139(33):11381–11391CrossRefGoogle Scholar
  30. 30.
    Durão P, Chen Z, Fernandes AT, Hildebrandt P, Murgida DH, Todorovic S, Pereira MM, Melo EP, Martins LO (2008) Copper incorporation into recombinant CotA laccase from Bacillus subtilis: characterization of fully copper loaded enzymes. J Biol Inorg Chem 13(2):183–193CrossRefGoogle Scholar
  31. 31.
    Webb SM, Dick GJ, Bargar JR, Tebo BM (2005) Evidence for the presence of Mn(III) intermediates in the bacterial oxidation of Mn(II). Proc Natl Acad Sci USA 102(15):5558–5563CrossRefGoogle Scholar
  32. 32.
    Archibald FS, Fridovich I (1982) The scavenging of superoxide radical by manganous complexes: in vitro. Arch Biochem Biophys 214(2):452–463CrossRefGoogle Scholar
  33. 33.
    Stoll S, Britt RD (2009) General and efficient simulation of pulse EPR spectra. Phys Chem Chem Phys 11(31):6614–6625CrossRefGoogle Scholar
  34. 34.
    Stoll S, Schweiger A (2006) EasySpin, a comprehensive software package for spectral simulation and analysis in EPR. J Magn Reson 178(1):42–55CrossRefGoogle Scholar
  35. 35.
    Campbell KA, Yikilmaz E, Grant CV, Gregory W, Miller A, Britt RD (1999) Parallel polarization EPR characterization of the Mn(III) center of oxidized manganese superoxide dismutase. J Am Chem Soc 121(19):4714–4715CrossRefGoogle Scholar
  36. 36.
    Campbell KA (1999) University of California Davis, Ph.D. dissertationGoogle Scholar
  37. 37.
    Hendrich MP, Debrunner PG (1989) Integer-spin electron paramagnetic resonance of iron proteins. Biophys J 56(3):489–506CrossRefGoogle Scholar
  38. 38.
    Münck E, Ksurerus K, Hendrich MP (1993) Combining Mössbauer spectroscopy with integer spin electron paramagnetic resonance. Methods Enzymol Acad Press 227:463–479CrossRefGoogle Scholar
  39. 39.
    Sharma A, Gaidamakova EK, Grichenko O, Matrosova VY, Hoeke V, Klimenkova P, Conze IH, Volpe RP, Tkavc R, Gostinčar C, Gunde-Cimerman N, DiRuggiero J, Shuryak I, Ozarowski A, Hoffman BM, Daly MJ (2017) Across the tree of life, radiation resistance is governed by antioxidant Mn2+, gauged by paramagnetic resonance. Proc Natl Acad Sci 114(44):E9253–E9260CrossRefGoogle Scholar
  40. 40.
    Krzystek J, Telser J, Pardi LA, Goldberg DP, Hoffman BM, Brunel L-C (1999) High-frequency and -field electron paramagnetic resonance of high-spin manganese(III) in porphyrinic complexes. Inorg Chem 38(26):6121–6129CrossRefGoogle Scholar
  41. 41.
    Goldberg DP, Telser J, Krzystek J, Montalban AG, Brunel L-C, Barrett AGM, Hoffman BM (1997) EPR spectra from “EPR-Silent” species: high-field EPR spectroscopy of manganese(III) porphyrins. J Am Chem Soc 119(37):8722–8723CrossRefGoogle Scholar
  42. 42.
    Anne-Laure B, Dante G, Roberta S, Luca AG, Andrea C, Fabretti AC, Uytterhoeven MG (1997) Electronic structure of Manganese(III) compounds from high-frequency EPR spectra. Angew Chem Int Ed Engl 36(21):2329–2331CrossRefGoogle Scholar
  43. 43.
    Telser J, Pardi LA, Krzystek J, Brunel L-C (1998) EPR spectra from “EPR-Silent” species: high-field EPR spectroscopy of aqueous chromium(II). Inorg Chem 37(22):5769–5775CrossRefGoogle Scholar
  44. 44.
    Campbell KA, Force DA, Nixon PJ, Dole F, Diner BA, Britt RD (2000) Dual-mode EPR detects the initial intermediate in photoassembly of the photosystem II Mn cluster: the influence of amino acid residue 170 of the D1 polypeptide on Mn coordination. J Am Chem Soc 122(15):3754–3761CrossRefGoogle Scholar
  45. 45.
    Whittaker JW, Whittaker MM (1991) Active site spectral studies on manganese superoxide dismuase. J Am Chem Soc 113(17):5528–5540CrossRefGoogle Scholar
  46. 46.
    Griffith JS (1971) The theory of transition-metal ions. Cambridge University Press, LondonGoogle Scholar
  47. 47.
    Sheng Y, Stich TA, Barnese K, Gralla EB, Cascio D, Britt RD, Cabelli DE, Valentine JS (2011) Comparison of two yeast MnSODs: mitochondrial Saccharomyces cerevisiae versus cytosolic Candida albicans. J Am Chem Soc 133(51):20878–20889CrossRefGoogle Scholar
  48. 48.
    Mossin S, Weihe H, Barra A-L (2002) Is the axial zero-field splitting parameter of tetragonally elongated high-spin Manganese(III) complexes always negative? J Am Chem Soc 124(30):8764–8765CrossRefGoogle Scholar
  49. 49.
    Campbell KA, Lashley MR, Wyatt JK, Nantz MH, Britt RD (2001) Dual-mode EPR study of Mn(III) salen and the Mn(III) salen-catalyzed epoxidation of cis-β-methylstyrene. J Am Chem Soc 123(24):5710–5719CrossRefGoogle Scholar
  50. 50.
    Zhu W, Wilcoxen J, Britt RD, Richards NGJ (2016) Formation of hexacoordinate Mn(III) in Bacillus subtilis oxalate decarboxylase requires catalytic turnover. Biochemistry 55(3):429–434CrossRefGoogle Scholar
  51. 51.
    Krivokapic I, Noble C, Klitgaard S, Tregenna-Piggott P, Weihe H, Barra AL (2005) Anisotropic hyperfine interaction in the manganese(III) hexaaqua ion. Angew Chem Int Ed 44(23):3613–3616CrossRefGoogle Scholar
  52. 52.
    Tregenna-Piggott PLW, Weihe H, Barra A-L (2003) High-field, multifrequency EPR study of the [Mn(OH2)6]3+ cation: influence of π-bonding on the ground state zero-field-splitting parameters. Inorg Chem 42(25):8504–8508CrossRefGoogle Scholar
  53. 53.
    Klewicki JK, Morgan JJ (1998) Kinetic behavior of Mn(III) complexes of pyrophosphate, EDTA, and citrate. Environ Sci Technol 32(19):2916–2922CrossRefGoogle Scholar
  54. 54.
    Gromov I, Marchesini A, Farver O, Pecht I, Goldfar D (1999) Azide binding to the trinuclear copper center in laccase and ascorbate oxidase. Eur J Biochem 266(3):820–830CrossRefGoogle Scholar
  55. 55.
    Soldatova AV, Butterfield C, Oyerinde OF, Tebo BM, Spiro TG (2012) Multicopper oxidase involvement in both Mn(II) and Mn(III) oxidation during bacterial formation of MnO2. J Biol Inorg Chem 17(8):1151–1158CrossRefGoogle Scholar
  56. 56.
    Dugad LB, Behere DV, Marathe VR, Mitra S (1984) Magnetic properties and electronic structure of manganese(III) porphyrins. Chem Phys Lett 104(4):353–356CrossRefGoogle Scholar
  57. 57.
    Kennedy BJ, Murray KS (1985) Magnetic properties and zero-field splitting in high-spin manganese(III) complexes. 2. Axially ligated manganese(III) porphyrin complexes. Inorg Chem 24(10):1557–1560CrossRefGoogle Scholar
  58. 58.
    White KN, Conesa C, Sánchez L, Amini M, Farnaud S, Lorvoralak C, Evans RW (2012) The transfer of iron between ceruloplasmin and transferrins. Biochim Biophys Acta Gen Subjects 1820(3):411–416CrossRefGoogle Scholar

Copyright information

© SBIC 2018

Authors and Affiliations

  • Lizhi Tao
    • 1
  • Troy A. Stich
    • 1
  • Alexandra V. Soldatova
    • 3
  • Bradley M. Tebo
    • 4
  • Thomas G. Spiro
    • 3
  • William H. Casey
    • 1
    • 2
  • R. David Britt
    • 1
    Email author
  1. 1.Department of ChemistryUniversity of CaliforniaDavisUSA
  2. 2.Department of GeologyUniversity of CaliforniaDavisUSA
  3. 3.Department of ChemistryUniversity of WashingtonSeattleUSA
  4. 4.Division of Environmental and Biomolecular SystemsOregon Health and Science UniversityPortlandUSA

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