Abstract
Proton Hyperfine Sublevel Correlation (1H-HYSCORE) experiments have been used to probe the ligation structure of the Fe(II) active site of taurine:2-oxoglutarate dioxygenase (TauD), a non-heme Fe(II) hydroxylase. To facilitate Electron Paramagnetic Resonance (EPR) experiments, Fe(II) derivatives of the enzyme were studied using nitric oxide as a substitute for molecular oxygen. The addition of NO to the enzyme yields an S = 3/2 {FeNO}7 paramagnetic center characterized by nearly axial EPR spectra with g⊥ = 4 and g||= 2. Using results from (i) an X-ray crystallographic study of TauD crystallized under anaerobic conditions in the presence of both cosubstrate 2-oxoglutarate and substrate taurine, (ii) a published theoretical description of the {FeNO}7 derivative of this form of the enzyme, and (iii) previous 2H-Electron Spin Echo Envelope Modulation (ESEEM) studies, we were able to assign the proton cross peaks detected in orientation-selected 1H-HYSCORE spectra. Discrete contributions from the protons of two coordinated histidine ligands were resolved. If substrate taurine is absent from the complex, orientation-selective HYSCORE spectra show cross peaks that are less resolved and when combined with information obtained from continuous wave EPR, support an alternate binding scheme for 2-oxoglutarate. HYSCORE studies of TauD in the absence of 2-oxoglutarate show additional 1H cross peaks that can be assigned to two distinct bound water molecules. In addition, 1H and 14N cross peaks that arise from the coordinated histidine side chains show a change in NO coordination for this species. For all of the TauD species, 1H hyperfine couplings and their orientations are sensitive to the detailed electronic structure of the {FeNO}7 center.
Similar content being viewed by others
Data availability
The data presented in this article and the MATLAB scripts used for data analysis can be made available upon request to the corresponding author.
References
E.G. Kovaleva, J.D. Lipscomb, Nat. Chem. Biol. 4, 186 (2008)
C.Q. Herr, R.P. Hausinger, Trends Biochem. Sci. 43, 517 (2018)
Y. Wang, J. Li, A. Liu, J. Biol. Inorg. Chem. 22, 395 (2017)
E.L. Hegg, L. Que, Eur. J. Biochem. 250, 625 (1997)
S. Kal, L. Que, J. Biol. Inorg. Chem. 22, 339 (2017)
J.C. Salerno, J.N. Siedow, Biochim. Biophys. Acta 579, 246 (1979)
J.H. Enemark, R.D. Feltham, Coord. Chem. Rev. 13, 339 (1974)
D.M. Arciero, J.D. Lipscomb, B.H. Huynh, T.A. Kent, E. Münck, J. Biol. Chem. 258, 14981 (1983)
V.J. Chen, A.M. Orville, M.R. Harpel, C.A. Frolik, K.K. Surerus, E. Münck, J.D. Lipscomb, J. Biol. Chem. 264, 21677 (1989)
A.M. Orville, V.J. Chen, A. Kriauciunas, M.R. Harpel, B.G. Fox, E. Münck, J.D. Lipscomb, Biochemistry 31, 4602 (1992)
D.M. Arciero, A.M. Orville, J.D. Lipscomb, J. Biol. Chem. 260, 14035 (1985)
D.M. Arciero, J.D. Lipscomb, J. Biol. Chem. 261, 2170 (1986)
B.M. Hoffman, J. Martinsen, R.A. Venters, J. Magn. Reson. 59, 110 (1984)
G.C. Hurst, T.A. Henderson, R.W. Kreilick, J. Am. Chem. Soc. 107, 7294 (1985)
T.C. Yang, M.D. Wolfe, M.B. Nebergall, Y. Mekmouche, J.D. Lipscomb, B.M. Hoffman, J. Am. Chem. Soc. 125, 7056 (2003)
A.M. Rocklin, D.L. Tierney, V. Kofman, N.M. Brunhuber, B.M. Hoffman, R.E. Christoffersen, N.O. Reich, J.D. Lipscomb, L. Que, Proc. Natl. Acad. Sci. USA 96, 7905 (1999)
D.L. Tierney, A.M. Rocklin, J.D. Lipscomb, L. Que, B.M. Hoffman, J. Am. Chem. Soc. 127, 7005 (2005)
T.M. Casey, P.K. Grzyska, R.P. Hausinger, J. McCracken, J. Phys. Chem. B 117, 10384 (2013)
R.J. Martinie, C.J. Pollock, M.L. Matthews, J.M. Bollinger, C. Krebs, A. Silakov, Inorg. Chem. 56, 13382 (2017)
R.J. Martinie, J. Livada, W. Chang, M.T. Green, C. Krebs, J.M. Bollinger, A. Silakov, J. Am. Chem. Soc. 137, 6912 (2015)
M.D. Krzyaniak, B.E. Eser, H.R. Ellis, P.F. Fitzpatrick, J. McCracken, Biochemistry 52, 8430 (2013)
C.A. Brown, M.A. Pavlosky, T.E. Westre, Y. Zhang, B. Hedman, K.O. Hodgson, E.I. Solomon, J. Am. Chem. Soc. 117, 715 (1995)
S. Ye, J.C. Price, E.W. Barr, M.T. Green, J.M. Bollinger, C. Krebs, F. Neese, J. Am. Chem. Soc. 132, 4739 (2010)
A. Bencini, D. Gatteschi, EPR of Exchange Coupled Systems (Springer, Berlin, 1990)
J. McCracken, B.E. Eser, D. Mannikko, M.D. Krzyaniak, P.F. Fitzpatrick, Biochemistry 54, 3759 (2015)
R.P. Hausinger, in 2-Oxoglutarate-Dependent Oxygenases. ed. by R.P. Hausinger, C.J. Schofield (Royal Society of Chemistry, London, 2015), pp. 1–58
H.M. Hanauske-Abel, V. Günzler, J. Theor. Biol. 94, 421 (1982)
J.M. Bollinger, W. Chang, M.L. Matthews, R.J. Martinie, A.K. Boal, C. Krebs, in 2-Oxoglutarate-Dependent Oxygenases. ed. by R.P. Hausinger, C.J. Schofield (Royal Society of Chemistry, London, 2015), pp. 95–123
J.M. Elkins, M.J. Ryle, I.J. Clifton, J.C.D. Hotopp, J.S. Lloyd, N.I. Burzlaff, J.E. Baldwin, R.P. Hausinger, P.L. Roach, Biochemistry 41, 5185 (2002)
J.J. Shane, P. Höfer, E.J. Reijerse, E. de Boer, J. Magn. Reson. 99, 596 (1992)
C. Gemperle, G. Aebli, A. Schweiger, R.R. Ernst, J. Magn. Reson. 88, 241 (1990)
S. Stoll, A. Schweiger, J. Magn. Reson. 178, 42 (2006)
C.D. Brown, M.L. Neidig, M.B. Neibergall, J.D. Lipscomb, E.I. Solomon, J. Am. Chem. Soc. 129, 7427 (2007)
S. Stoll, R.D. Britt, Phys. Chem. Chem. Phys. 11, 6614 (2009)
F. Aquino, J.H. Rodriguez, J. Phys. Chem. A 113, 9150 (2009)
A. Schweiger, G. Jeschke, Principles of Pulse Electron Paramagnetic Resonance (Oxford University Press, Oxford, 2001)
A. Pöppl, L. Kevan, J. Phys. Chem. 100, 3387 (1996)
J. McCracken, P.J. Cappillino, J.S. McNally, M. Howart, M.D. Krzyaniak, J.P. Caradonna, Inorg. Chem. 54, 6486 (2015)
K.M. Davis, M. Altmyer, R.J. Martinie, I. Schaperdoth, C. Krebs, J.M. Bollinger, A.K. Boal, Biochemistry 58, 4218 (2019)
Acknowledgements
We thank Meng Li who initiated these studies. J.M. would like to thank Drs. Gareth and Sandra Eaton for their support over the past 35 years.
Funding
This work was supported by National Institutes of Health grants GM065384 (R.P.H.), GM054065 (J.M.) and RR15880 (J.M.).
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
McCracken, J., Casey, T.M. & Hausinger, R.P. 1H-HYSCORE Reveals Structural Details at the Fe(II) Active Site of Taurine:2-Oxoglutarate Dioxygenase. Appl Magn Reson 52, 971–994 (2021). https://doi.org/10.1007/s00723-020-01288-w
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00723-020-01288-w