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Information on EC 1.6.2.4 - NADPH-hemoprotein reductase and Organism(s) Priestia megaterium and UniProt Accession P14779

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EC Tree
     1 Oxidoreductases
         1.6 Acting on NADH or NADPH
             1.6.2 With a heme protein as acceptor
                1.6.2.4 NADPH-hemoprotein reductase
IUBMB Comments
A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases (e.g. EC 1.14.14.1, unspecific monooxygenase) by stabilizing the one-electron reduced form of the flavin cofactors FAD and FMN. It also reduces cytochrome b5 and cytochrome c. The number n in the equation is 1 if the hemoprotein undergoes a 2-electron reduction, and is 2 if it undergoes a 1-electron reduction.
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Priestia megaterium
UNIPROT: P14779
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Word Map
The taxonomic range for the selected organisms is: Priestia megaterium
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Synonyms
nadph-cytochrome p-450 reductase, cytochrome p450 reductase, p450 reductase, p-450 reductase, nadph-p450 reductase, p450 bm3, p450 oxidoreductase, nadph cytochrome p450 reductase, cytochrome p450 oxidoreductase, cypor, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
flavocytochrome P450 BM3
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P450 BM3
bifunctional P-450/NADPH-P450 reductase, EC1.14.14.1/EC1.6.2.4
aldehyde reductase (NADPH-dependent)
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-
-
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cytochrome c reductase (reduced nicotinamide adenine dinucleotide phosphate, NADPH, NADPH-dependent)
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-
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cytochrome P450 reductase
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dihydroxynicotinamide adenine dinucleotide phosphate-cytochrome c reductase
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-
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FAD-cytochrome c reductase
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-
-
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ferrihemprotein P450 reductase
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-
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NADP-cytochrome c reductase
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-
-
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NADP-cytochrome reductase
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-
-
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NADPH-cytochrome c oxidoreductase
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-
-
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NADPH-cytochrome c reductase
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-
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NADPH-cytochrome p-450 reductase
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-
-
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NADPH-cytochrome P450 (CYP) oxidoreductase
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-
-
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NADPH-dependent cytochrome c reductase
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-
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NADPH-ferricytochrome c oxidoreductase
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-
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P450R
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-
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reduced nicotinamide adenine dinucleotide phosphate-cytochrome c reductase
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-
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reductase, cytochrome c (reduced nicotinamide adenine dinucleotide phosphate)
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-
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TPNH-cytochrome c reductase
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-
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TPNH2 cytochrome c reductase
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-
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-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
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-
-
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oxidation
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-
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reduction
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-
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SYSTEMATIC NAME
IUBMB Comments
NADPH:hemoprotein oxidoreductase
A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases (e.g. EC 1.14.14.1, unspecific monooxygenase) by stabilizing the one-electron reduced form of the flavin cofactors FAD and FMN. It also reduces cytochrome b5 and cytochrome c. The number n in the equation is 1 if the hemoprotein undergoes a 2-electron reduction, and is 2 if it undergoes a 1-electron reduction.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-03-4
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
show the reaction diagram
ferricytochrome c + NADPH + H+
ferrocytochrome c + NADP+
show the reaction diagram
-
-
-
-
r
NADPH + ferricytochrome P450
ferrocytochrome P450 + NADP+
show the reaction diagram
-
during hydroxylation of fatty acids through the bacterial fusion protein P450BM3
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-
?
NADPH + H+ + ferricytochrome c
NADP+ + ferrocytochrome c
show the reaction diagram
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-
-
-
?
NADPH + H+ + oxidized CYP106A1
NADP+ + reduced CYP106A1
show the reaction diagram
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-
-
-
?
NADPH + H+ + oxidized CYP21A2
NADP+ + reduced CYP21A2
show the reaction diagram
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-
-
-
?
additional information
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
NADPH + ferricytochrome P450
ferrocytochrome P450 + NADP+
show the reaction diagram
-
during hydroxylation of fatty acids through the bacterial fusion protein P450BM3
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-
?
additional information
?
-
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part of the bacterial fusion protein P450BM3 composed of cytochrome P450 fatty acid hydroxylase and NADPH cytochrome P450 reductase
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-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Iron
investigation of the haem-domain
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-phenylimidazole
inhibitory azole ligand that co-ordinates the haem iron in the sixth (axial) position
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CO
-
stimulation
hexadecanol
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stimulation
laurate
palmitate
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stimulation
Tetradecanol
-
stimulation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0167
ferricytochrome c
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at pH 7.5 and 25°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
43.03 - 417
ferricytochrome c
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
expression in Escherichia coli
SwissProt
Manually annotated by BRENDA team
PDB
SCOP
CATH
UNIPROT
ORGANISM
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
sitting-drop technique, 4°C, A264Q crystals are obtained with a mother liquor of 100 mM cacodylic acid (pH 6.0), 160 mM MgCl2 and 16% PEG 3350. A264M crystals are obtained with mother liquor of 100 mM cacodylic acid (pH 6.0), 140 mM MgCl2 and 18% PEG 3350. A264C crystals are obtained under the same conditions, but with 100 mM MgCl2. Crystals are flash-frozen in liquid nitrogen using 10% PEG 200 as cryoprotectant. Crystal structures of the mutant haem domains demonstrate axial ligation of P450 haem iron by methionine and glutamine ligands trans to the cysteine thiolate, creating novel haem iron ligand sets in the A264M/Q variants
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A264C
no notable decreases in expression of the respective haem domains or flavocytochromes. For A264C haem domain, fatty-acid-induced perturbation of the spinstate equilibrium is observed in some cases, but the extent of high-spin conversion is substantially less than observed for wild-type with arachidonate
A264M
higher dodecanoate affinity than wild-type BM3. For the A264M mutant, a more substantial fatty-acid-induced high-spin conversion of haem iron is observed than for A264C, albeit slightly less than for wild-type with the same fatty acids.
A264Q
no notable decreases in expression of the respective haem domains or flavocytochromes
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
54
-
the reductase loses 50% of its activity after 10 min at 54°C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
expressed in E. coli, -80°C
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
homogeneity, chromatographic methods (DEAE-Sepharose, Q-Sepharose and hydroxyapatite resins)
expressed in Escherichia coli, column chromatography on 2',5'-ADP-agarose
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Ni-NTA column chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expressed in Escherichia coli C43(DE3) cells
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expression in Escherichia coli
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
oxidation of the reduced P450BM3 by cytochrome c, ferricyanide or 2,6-dichlorophenolindophenol rapidly restores electron transfer and hydroxylase activity
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Murataliev, M.B.; Feyereisen, R.
Functional interactions in cytochrome P450BM3. Fatty acid substrate binding alters electron-transfer properties of the flavoprotein domain
Biochemistry
35
15029-15037
1996
Priestia megaterium
Manually annotated by BRENDA team
Girvan, H.M.; Toogood, H.; Littleford, R.E.; Seward, H.E.; Smith, W.E.; Ekanem, I.; Leys, D.; Cheesman, M.; Munro, A.W.
Novel heme coordination variants of flavocytochrome P450 BM3
Biochem. J.
417
65-76
2009
Priestia megaterium (P14779), Priestia megaterium
Manually annotated by BRENDA team
Milhim, M.; Gerber, A.; Neunzig, J.; Hannemann, F.; Bernhardt, R.
A novel NADPH-dependent flavoprotein reductase from Bacillus megaterium acts as an efficient cytochrome P450 reductase
J. Biotechnol.
231
83-94
2016
Priestia megaterium, Priestia megaterium DSM319
Manually annotated by BRENDA team