Cited by 19Lipid Membrane Structure and BehaviorPharmacogenetics and Drug MetabolismMetal-Catalyzed Oxygenation Mechanisms

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Structural and Functional Studies of the Membrane-Binding Domain of NADPH-Cytochrome P450 Oxidoreductase

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Xia, Chuanwu, Shen, Anna L., Duangkaew, Panida, Kotewong, Rattanawadee, Rongnoparut, Pornpimol, Feix, Jimmy, Kim, Jung-Ja P.·2019·Biochemistry

DOI 10.1021/acs.biochem.9b00130PMC PMC6873807

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Abstract

NADPH-cytochrome P450 oxidoreductase (CYPOR), the essential flavoprotein of the microsomal cytochrome P450 monooxygenase system, is anchored in the phospholipid bilayer by its amino-terminal membrane-binding domain (MBD), which is necessary for efficient electron transfer to cytochrome P450. Although crystallographic and kinetic studies have established the structure of the soluble catalytic domain and the role of conformational motions in the control of electron transfer, the role of the MBD is largely unknown. We examined the role of the MBD in P450 catalysis through studies of amino-terminal deletion mutants and site-directed spin labeling. We show that the MBD spans the membrane and present a model for the orientation of CYPOR on the membrane capable of forming a complex with cytochrome P450. EPR power saturation measurements of CYPOR mutants in liposomes containing a lipid/Ni(II) chelate identified a region of the soluble domain interacting with the membrane. The deletion of more than 29 residues from the N-terminus of CYPOR decreases cytochrome P450 activity concomitant with alterations in electrophoretic mobility and an increased resistance to protease digestion. The altered kinetic properties of these mutants are consistent with electron transfer through random collisions rather than via formation of a stable CYPOR-P450 complex. Purified MBD binds weakly to cytochrome P450, suggesting that other interactions are also required for CYPOR-P450 complex formation. We propose that the MBD and flexible tether region of CYPOR, residues 51-63, play an important role in facilitating the movement of the soluble domain relative to the membrane and in promoting multiple orientations that permit specific interactions of CYPOR with its varied partners.

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Hepatic Triphosphopyridine Nucleotide-Cytochrome c Reductase: Isolation, Characterization, and Kinetic Studies
Phillips, A. H., & LANGDON, R. G. (1962) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)73803-4 · Cited by 1,424
Three-dimensional structure of NADPH-cytochrome P450 reductase: prototype for FMN- and FAD-containing enzymes
Wang, M., Roberts, D. L., Paschke, R., Shea, T. M., Masters, B. S. S., & Kim, J.-J. P. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.16.8411 · Cited by 744
Self-citation
Role of Hemoprotein P-450 in Fatty Acid ω-Hydroxylation in a Soluble Enzyme System from Liver Microsomes
Lu, A., & Coon, M. J. (1968) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)56992-7 · Cited by 627
Recent advances in site-directed spin labeling of proteins
Hubbell, W. L., Gross, A., Langen, R., & Lietzow, M. A. (1998) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(98)80158-9 · Cited by 560
Structure of a cytochrome P450-redox partner electron-transfer complex
Sevrioukova, I. F., Li, H., Zhang, H., Peterson, J. A., & Poulos, T. L. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.5.1863 · Cited by 501
A collision gradient method to determine the immersiondepth of nitroxides in lipid bilayers: application to spin-labeled mutants ofbacteriorhodopsin.
Altenbach, C., Greenhalgh, D. A., Khorana, H. G., & Hubbell, W. L. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.5.1667 · Cited by 411
Structure of Mammalian Cytochrome P450 2B4 Complexed with 4-(4-Chlorophenyl)imidazole at 1.9-Å Resolution
Scott, E. E., White, M. A., He, Y. A., Johnson, E. F., Stout, C. D., & Halpert, J. R. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m403349200 · Cited by 282
Structural analysis of the FMN binding domain of NADPH-cytochrome P-450 oxidoreductase by site-directed mutagenesis
Shen, A. L., Porter, T. D., Wilson, T. E., & Kasper, C. B. (1989) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)83274-4 · Cited by 264
Self-citation
Phenobarbital mechanistic data and risk assessment: Enzyme induction, enhanced cell proliferation, and tumor promotion
Whysner, J. A. (1996) · Pharmacology & Therapeutics · doi:10.1016/0163-7258(96)00067-8 · Cited by 233
Purification of cytochrome P-450, NADPH-cytochrome P-450 reductase, and epoxide hydratase from a single preparation of rat liver microsomes
Guengerich, F. P., & Martin, M. V. (1980) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(80)90119-8 · Cited by 228
Interactions of cytochrome P-450, NADPH-cytochrome P-450 reductase, phospholipid, and substrate in the reconstituted liver microsomal enzyme system.
French, J. S., Guengerich, F. P., & Coon, M. J. (1980) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)85640-5 · Cited by 226
Structure and Function of an NADPH-Cytochrome P450 Oxidoreductase in an Open Conformation Capable of Reducing Cytochrome P450
Hamdane, D., Xia, C., Im, S., Zhang, H., Kim, J. P., & Waskell, L. A. (2009) · Journal of Biological Chemistry · doi:10.1074/jbc.m807868200 · Cited by 225
Self-citation
NADPH-Cytochrome P450 Oxidoreductase
Hubbard, P. A., Shen, A. L., Paschke, R., Kasper, C. B., & Kim, J. P. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m101731200 · Cited by 220
Self-citation
Studies on the Mechanism of Microsomal Triphosphopyridine Nucleotide-Cytochrome c Reductase
Masters, B. S. S., Kamin, H., Gibson, Q. H., & Williams, C. H. (1965) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)45262-8 · Cited by 202
Methods and Applications of Site-Directed Spin Labeling EPR Spectroscopy
Klug, C. S., & Feix, J. B. (2007) · Methods in cell biology · doi:10.1016/s0091-679x(07)84020-9 · Cited by 194
Self-citation
Properties of NADPH-cytochrome P-450 reductase purified from rabbit liver microsomes
French, J. S., & Coon, M. J. (1979) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(79)90383-7 · Cited by 180
Structural features of liver microsomal NADPH-cytochrome P-450 reductase. Hydrophobic domain, hydrophilic domain, and connecting region.
Black, S. D., & Coon, M. J. (1982) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)83868-1 · Cited by 174
[29] Assay of formaldehyde generated during microsomal oxidation reactions
Werringloer, J. (1978) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(78)52031-4 · Cited by 157
Role of Acidic Residues in the Interaction of NADPH-Cytochrome P450 Oxidoreductase with Cytochrome P450 and Cytochrome c
Shen, A. L., & Kasper, C. B. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.46.27475 · Cited by 135
Self-citation
Studies on the association of cytochrome P-450 and NADPH-cytochrome c reductase during catalysis in a reconstituted hydroxylating system.
Miwa, G. T., West, S. B., Huang, M., & Lu, A. (1979) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)50470-1 · Cited by 134
Conformation of spin-labeled melittin at membrane surfaces investigated by pulse saturation recovery and continuous wave power saturation electron paramagnetic resonance
Altenbach, C., Froncisz, W., Hyde, J. S., & Hubbell, W. L. (1989) · Biophysical Journal · doi:10.1016/s0006-3495(89)82765-1 · Cited by 133
Molecular mechanisms of the microsomal mixed function oxidases and biological and pathological implications
Cederbaum, A. I. (2014) · Redox Biology · doi:10.1016/j.redox.2014.11.008 · Cited by 132
Role of a hydrophobic polypeptide in the N-terminal region of NADPH-cytochrome P-450 reductase in complex formation with P-450lm
Black, S. D., French, J. S., Williams, C. H., & Coon, M. J. (1979) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(79)91238-5 · Cited by 115
Structural basis for human NADPH-cytochrome P450 oxidoreductase deficiency
Xia, C., Panda, S. P., Marohnic, C. C., Pavel Martasek, E., Masters, B. S. S., & Kim, J. P. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1106632108 · Cited by 115
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Xia, C., Hamdane, D., Shen, A. L., Choi, V. W., Kasper, C. B., Pearl, N. M., Zhang, H., Im, S., Waskell, L. A., & Kim, J. P. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m111.230532 · Cited by 112
Self-citation
Modulation of the Cytochrome P450 Reductase Redox Potential by the Phospholipid Bilayer
Das, A., & Sligar, S. G. (2009) · Biochemistry · doi:10.1021/bi9011435 · Cited by 95
Interactions among P450 Enzymes When Combined in Reconstituted Systems: Formation of a 2B4−1A2 Complex with a High Affinity for NADPH−Cytochrome P450 Reductase
Backes, W. L., Batie, C. J., & Cawley, G. F. (1998) · Biochemistry · doi:10.1021/bi980674a · Cited by 87
Cytochrome P450 Organization and Function Are Modulated by Endoplasmic Reticulum Phospholipid Heterogeneity
Brignac‐Huber, L. M., Park, J. W., Reed, J. R., & Backes, W. L. (2016) · Drug Metabolism and Disposition · doi:10.1124/dmd.115.068981 · Cited by 68
Complete amino acid sequence of NADPH-cytochrome P-450 reductase from porcine hepatic microsomes
Haniu, M., Iyanagi, T., Miller, P., Lee, T. D., & Shively, J. E. (1986) · Biochemistry · doi:10.1021/bi00372a018 · Cited by 64
The N-Terminal Membrane Domain of Yeast NADPH-Cytochrome P450 (CYP) Oxidoreductase Is Not Required for Catalytic Activity in Sterol Biosynthesis or in Reconstitution of CYP Activity
Kanamarlapudi, V., Lamb, D. C., Kelly, D. E., Manning, N. J., & Kelly, S. L. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.8.4492 · Cited by 58
Mechanistic Studies on the Reductive Half-reaction of NADPH-Cytochrome P450 Oxidoreductase
Shen, A. L., Sem, D. S., & Kasper, C. B. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.9.5391 · Cited by 58
Self-citation
Isolation of the membrane-binding peptide of NADPH-cytochrome P-450 reductase. Characterization of the peptide and its role in the interaction of reductase with cytochrome P-450.
Gum, J. R., & Strobel, H. W. (1981) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)68987-8 · Cited by 56
Mapping of the Residues Involved in a Proposed β-Strand Located in the Ferric Enterobactin Receptor FepA Using Site-Directed Spin-Labeling
Klug, C. S., Su, W., & Feix, J. B. (1997) · Biochemistry · doi:10.1021/bi971232m · Cited by 55
Self-citation
Identification and characterization of an NADPH-cytochrome P450 reductase derived peptide involved in binding to cytochrome P450
Nadler, S. G., & Strobel, H. W. (1991) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(91)90542-q · Cited by 55
Membrane environment drives cytochrome P450's spin transition and its interaction with cytochrome b 5
Ravula, T., Barnaba, C., Mahajan, M. K., Anantharamaiah, G. M., Im, S., Waskell, L. A., & Ramamoorthy, A. (2017) · Chemical Communications · doi:10.1039/c7cc07520k · Cited by 42
Probing the Transmembrane Structure and Dynamics of Microsomal NADPH-cytochrome P450 oxidoreductase by Solid-State NMR
Huang, R., Yamamoto, K., Zhang, M., Popovych, N., Hung, I., Im, S., Gan, Z., Waskell, L. A., & Ramamoorthy, A. (2014) · Biophysical Journal · doi:10.1016/j.bpj.2014.03.051 · Cited by 40
Identification of Protein Side Chains near the Membrane−Aqueous Interface: A Site-Directed Spin Labeling Study of KcsA
Gross, A., & Hubbell, W. L. (2001) · Biochemistry · doi:10.1021/bi015828s · Cited by 40
Differential Contributions of NADPH-Cytochrome P450 Oxidoreductase FAD Binding Site Residues to Flavin Binding and Catalysis
Shen, A. L., & Kasper, C. B. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m008380200 · Cited by 40
Self-citation
Membrane Topology of NADPH–Cytochrome P450 Reductase on the Endoplasmic Reticulum
Kida, Y., Ohgiya, S., Mihara, K., & Sakaguchi, M. (1998) · Archives of Biochemistry and Biophysics · doi:10.1006/abbi.1997.0553 · Cited by 39
Transmembrane Interactions of Full-length Mammalian Bitopic Cytochrome-P450-Cytochrome-b5 Complex in Lipid Bilayers Revealed by Sensitivity-Enhanced Dynamic Nuclear Polarization Solid-state NMR Spectroscopy
Yamamoto, K., Caporini, M. A., Im, S., Waskell, L. A., & Ramamoorthy, A. (2017) · Scientific Reports · doi:10.1038/s41598-017-04219-1 · Cited by 36
Interactions of cytochrome P450 2B4 with NADPH-cytochrome P450 reductase studied by fluorescent probe
Davydov, D. R., Knyushko, T. V., Kanaeva, I. P., Koen, Y. M., Samenkova, N. F., Archakov, A. I., & Hoa, G. H. B. (1996) · Biochimie · doi:10.1016/s0300-9084(97)82531-x · Cited by 32
Global Analysis of Protein−Protein Interactions Reveals Multiple CYP2E1−Reductase Complexes
Jamakhandi, A. P., Kuzmič, P., Sanders, D., & Miller, G. P. (2007) · Biochemistry · doi:10.1021/bi7003476 · Cited by 32
Binding of a physiological substrate causes large-scale conformational reorganization in cytochrome P450 51
Hargrove, T. Y., Wawrzak, Z., Fisher, P. M., Child, S. A., Nes, W. D., Guengerich, F. P., Waterman, M. R., & Lepesheva, G. I. (2018) · Journal of Biological Chemistry · doi:10.1074/jbc.ra118.005850 · Cited by 24
Beta sheet 2–alpha helix C loop of cytochrome P450 reductase serves as a docking site for redox partners
Jang, H.-H., Jamakhandi, A. P., Sullivan, S. Z., Yun, C., Hollenberg, P. F., & Miller, G. P. (2010) · Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics · doi:10.1016/j.bbapap.2010.02.003 · Cited by 23
Combined use of mass spectrometry and heterologous expression for identification of membrane-interacting peptides in cytochrome P450 46A1 and NADPH-cytochrome P450 oxidoreductase
Mast, N., Liao, W.-L., Pikuleva, I. A., & Turko, I. V. (2009) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2009.01.002 · Cited by 22
An Enzymatically Active Chimeric Protein Containing the Hydrophilic Form of NADPH-Cytochrome P450 Reductase Fused to the Membrane-Binding Domain of Cytochrome b5
Gilep, A. A., Guryev, O. L., Usanov, S. A., & Estabrook, R. W. (2001) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.2001.5075 · Cited by 21
An optical biosensor study of the interaction parameters and role of hydrophobic tails of cytochrome P450 2B4, b5 and NADPH‐flavoprotein in complex formation
Ivanov, Y. D., Kanaeva, I. P., Eldarov, M. A., Sklyabin, K. G., Lehnerer, M., Schulze, J., Hlavica, P., & Archakov, A. I. (1997) · IUBMB Life · doi:10.1080/15216549700203161 · Cited by 18
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Liu, K., Hughes, J. M. X., Hay, S., & Scrutton, N. S. (2017) · FEBS Journal · doi:10.1111/febs.14129 · Cited by 17
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See all 19

Authors: Xia, C., Shen, A. L., Duangkaew, P., Kotewong, R., Rongnoparut, P., Feix, J., & Kim, J.-J. P.
Affiliations: Medical College of Wisconsin (US)
University of Wisconsin–Madison (US)
Mahidol University (TH)
Corresponding author: Kim, Jung-Ja P.
ORCID: Xia, Chuanwu
Shen, Anna L.
Duangkaew, Panida
Rongnoparut, Pornpimol
Feix, Jimmy
Kim, Jung-Ja P.
Venue: Biochemistry
Publisher: American Chemical Society (ACS)
Published: 2019-04-22
Volume: 58
Issue: 19
Pages: 2408-2418
ISSN: 0006-2960 (ISSN-L)
1520-4995
DOI: 10.1021/acs.biochem.9b00130
PMC: PMC6873807
OpenAlex: W2940562984
Semantic Scholar: 40fce275fb8b489bdf467bb125854da271ef3a27
Cited by: 19
References: 51
Citations by year: 2026: 4 · 2025: 3 · 2024: 3 · 2022: 5 · 2021: 4
FWCI: 0.74
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: doi.org/10.15223/policy-029 (stm-asf)
doi.org/10.15223/policy-037 (stm-asf)
doi.org/10.15223/policy-045 (stm-asf)
Locations: Biochemistry (publishedVersion)
PubMed (publishedVersion)
PubMed Central (acceptedVersion)
Funders: National Institute of General Medical Sciences (GM114234, GM097031)
Thailand Research Fund
National Cancer Institute (CA22484)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
Keywords: NADPH-cytochrome P450 reductase, membrane-binding domain, cytochrome P450, electron transfer, flavoprotein, membrane anchoring, site-directed spin labeling, protein orientation in membranes
MeSH terms: Cell Membrane, Cysteine, Cytochrome P-450 Enzyme System, Electron Transport, Escherichia coli, Flavoproteins, Kinetics, Lipid Bilayers, Liposomes, NADP, NADPH-Ferrihemoprotein Reductase, Oxidoreductases, N-Demethylating, Plasmids, Protein Structure, Tertiary, Crystallography, X-Ray, Catalytic Domain, Sequence Analysis, Protein, Hydrophobic and Hydrophilic Interactions

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Structural and Functional Studies of the Membrane-Binding Domain of NADPH-Cytochrome P450 Oxidoreductase

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