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Protein

Bifunctional cytochrome P450/NADPH--P450 reductase 1

Gene

cypD

Organism
Bacillus subtilis (strain 168)
Status
Reviewed-Annotation score: -Experimental evidence at protein leveli

Functioni

Functions as a fatty acid monooxygenase. Catalyzes hydroxylation of a range of long-chain fatty acids, with a preference for long-chain unsaturated and branched-chain fatty acids over saturated fatty acids. Hydroxylation of myristic acid occurs mainly at the omega-2 position. Also displays a NADPH-dependent reductase activity in the C-terminal domain, which allows electron transfer from NADPH to the heme iron of the cytochrome P450 N-terminal domain (PubMed:15375636, PubMed:15122913). Is also able to catalyze efficient oxidation of sodium dodecyl sulfate (SDS) (PubMed:21048857).3 Publications

Catalytic activityi

RH + [reduced NADPH--hemoprotein reductase] + O2 = ROH + [oxidized NADPH--hemoprotein reductase] + H2O.2 Publications
NADPH + n oxidized hemoprotein = NADP+ + n reduced hemoprotein.2 Publications

Cofactori

Protein has several cofactor binding sites:

Kineticsi

kcat is 430 min(-1) for stearic acid hydroxylation. kcat is 5430 min(-1) for phytanic acid hydroxylation. kcat is 6105 min(-1) for 15-methylpalmitic acid hydroxylation. kcat is 11400 min(-1) for the reduction of cytochrome c. kcat is 38150 min(-1) for the reduction of ferricyanide (PubMed:15122913). kcat is 2244 min(-1) for oleic acid hydroxylation (PubMed:15375636).2 Publications
  1. KM=38.8 µM for stearic acid1 Publication
  2. KM=150 µM for phytanic acid1 Publication
  3. KM=56.8 µM for 15-methylpalmitic acid1 Publication
  4. KM=3.6 µM for NADPH1 Publication
  5. KM=17.9 mM for NADH1 Publication
  6. KM=6.9 µM for cytochrome c (in the reductase assay)1 Publication
  7. KM=153.4 µM for ferricyanide (in the reductase assay)1 Publication
  8. KM=17.36 µM for oleic acid1 Publication

    pH dependencei

    Optimum pH is 7.0.1 Publication

    Temperature dependencei

    Optimum temperature is 51 degrees Celsius. However, enzyme stability is dramatically reduced at this temperature, incubation for 30 minutes at 31 and 49 degrees Celsius results in 61% and 17% residual activity, respectively. Incubation at 60 degrees Celsius leads to total inactivation of the enzyme.1 Publication

    Sites

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Sitei271Important for catalytic activityBy similarity1
    Metal bindingi403Iron (heme axial ligand)By similarity1

    Regions

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Nucleotide bindingi499 – 504FMNBy similarity6
    Nucleotide bindingi546 – 549FMNBy similarity4
    Nucleotide bindingi580 – 582FMNBy similarity3
    Nucleotide bindingi588 – 590FMNBy similarity3

    GO - Molecular functioni

    • aromatase activity Source: UniProtKB-EC
    • fatty acid binding Source: UniProtKB
    • flavin adenine dinucleotide binding Source: UniProtKB
    • FMN binding Source: UniProtKB
    • heme binding Source: UniProtKB
    • iron ion binding Source: UniProtKB
    • NADPH-hemoprotein reductase activity Source: UniProtKB
    • oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen Source: UniProtKB

    GO - Biological processi

    • fatty acid oxidation Source: UniProtKB

    Keywordsi

    Molecular functionMonooxygenase, Multifunctional enzyme, Oxidoreductase
    Biological processElectron transport, Transport
    LigandFAD, Flavoprotein, FMN, Heme, Iron, Metal-binding, NADP

    Enzyme and pathway databases

    BioCyciBSUB:BSU07250-MONOMER
    SABIO-RKiO08394

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Bifunctional cytochrome P450/NADPH--P450 reductase 1Curated
    Alternative name(s):
    CYP102A21 Publication
    Fatty acid hydroxylase CypDCurated
    Flavocytochrome P450 102A2Curated
    Including the following 2 domains:
    Cytochrome P450 102A2Curated (EC:1.14.14.12 Publications)
    NADPH--cytochrome P450 reductaseCurated (EC:1.6.2.42 Publications)
    Gene namesi
    Name:cypDCombined sources
    Synonyms:cyp102A22 Publications, yetO1 Publication, yfnJ
    Ordered Locus Names:BSU07250
    OrganismiBacillus subtilis (strain 168)
    Taxonomic identifieri224308 [NCBI]
    Taxonomic lineageiBacteriaFirmicutesBacilliBacillalesBacillaceaeBacillus
    Proteomesi
    • UP000001570 Componenti: Chromosome

    Subcellular locationi

    GO - Cellular componenti

    Keywords - Cellular componenti

    Cytoplasm

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Mutagenesisi15P → S: Exhibits modified substrate specificity. Shows approximately 6- to 9-fold increased activity with SDS, lauric acid and 1,4-naphthoquinone, and enhanced activity for other substrates such as ethacrynic acid and epsilon-amino-n-caproic acid. 1 Publication1

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    ChainiPRO_00000522061 – 1061Bifunctional cytochrome P450/NADPH--P450 reductase 1Add BLAST1061

    Proteomic databases

    PaxDbiO08394
    PRIDEiO08394

    Interactioni

    Protein-protein interaction databases

    STRINGi224308.Bsubs1_010100004068

    Structurei

    3D structure databases

    ProteinModelPortaliO08394
    SMRiO08394
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Domaini493 – 632Flavodoxin-likePROSITE-ProRule annotationAdd BLAST140
    Domaini671 – 904FAD-binding FR-typePROSITE-ProRule annotationAdd BLAST234

    Region

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Regioni1 – 475Cytochrome P4501 PublicationAdd BLAST475
    Regioni476 – 1061NADPH--P450 reductase1 PublicationAdd BLAST586

    Sequence similaritiesi

    In the N-terminal section; belongs to the cytochrome P450 family.Curated

    Phylogenomic databases

    eggNOGiENOG4107EER Bacteria
    COG0369 LUCA
    COG2124 LUCA
    HOGENOMiHOG000093545
    InParanoidiO08394
    KOiK14338
    OMAiCEIRFER
    PhylomeDBiO08394

    Family and domain databases

    Gene3Di1.10.630.10, 1 hit
    1.20.990.10, 1 hit
    3.40.50.360, 1 hit
    InterProiView protein in InterPro
    IPR023206 Bifunctional_P450_P450_red
    IPR001128 Cyt_P450
    IPR017972 Cyt_P450_CS
    IPR036396 Cyt_P450_sf
    IPR003097 FAD-binding_1
    IPR017927 Fd_Rdtase_FAD-bd
    IPR001094 Flavdoxin-like
    IPR008254 Flavodoxin/NO_synth
    IPR001709 Flavoprot_Pyr_Nucl_cyt_Rdtase
    IPR029039 Flavoprotein-like_sf
    IPR023173 NADPH_Cyt_P450_Rdtase_alpha
    IPR001433 OxRdtase_FAD/NAD-bd
    IPR017938 Riboflavin_synthase-like_b-brl
    PfamiView protein in Pfam
    PF00667 FAD_binding_1, 1 hit
    PF00258 Flavodoxin_1, 1 hit
    PF00175 NAD_binding_1, 1 hit
    PF00067 p450, 1 hit
    PIRSFiPIRSF000209 Bifunctional_P450_P450R, 1 hit
    PRINTSiPR00369 FLAVODOXIN
    PR00371 FPNCR
    SUPFAMiSSF48264 SSF48264, 1 hit
    SSF52218 SSF52218, 1 hit
    SSF63380 SSF63380, 1 hit
    PROSITEiView protein in PROSITE
    PS00086 CYTOCHROME_P450, 1 hit
    PS51384 FAD_FR, 1 hit
    PS50902 FLAVODOXIN_LIKE, 1 hit

    Sequencei

    Sequence statusi: Complete.

    O08394-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MKETSPIPQP KTFGPLGNLP LIDKDKPTLS LIKLAEEQGP IFQIHTPAGT
    60 70 80 90 100
    TIVVSGHELV KEVCDEERFD KSIEGALEKV RAFSGDGLFT SWTHEPNWRK
    110 120 130 140 150
    AHNILMPTFS QRAMKDYHEK MVDIAVQLIQ KWARLNPNEA VDVPGDMTRL
    160 170 180 190 200
    TLDTIGLCGF NYRFNSYYRE TPHPFINSMV RALDEAMHQM QRLDVQDKLM
    210 220 230 240 250
    VRTKRQFRYD IQTMFSLVDS IIAERRANGD QDEKDLLARM LNVEDPETGE
    260 270 280 290 300
    KLDDENIRFQ IITFLIAGHE TTSGLLSFAT YFLLKHPDKL KKAYEEVDRV
    310 320 330 340 350
    LTDAAPTYKQ VLELTYIRMI LNESLRLWPT APAFSLYPKE DTVIGGKFPI
    360 370 380 390 400
    TTNDRISVLI PQLHRDRDAW GKDAEEFRPE RFEHQDQVPH HAYKPFGNGQ
    410 420 430 440 450
    RACIGMQFAL HEATLVLGMI LKYFTLIDHE NYELDIKQTL TLKPGDFHIS
    460 470 480 490 500
    VQSRHQEAIH ADVQAAEKAA PDEQKEKTEA KGASVIGLNN RPLLVLYGSD
    510 520 530 540 550
    TGTAEGVARE LADTASLHGV RTKTAPLNDR IGKLPKEGAV VIVTSSYNGK
    560 570 580 590 600
    PPSNAGQFVQ WLQEIKPGEL EGVHYAVFGC GDHNWASTYQ YVPRFIDEQL
    610 620 630 640 650
    AEKGATRFSA RGEGDVSGDF EGQLDEWKKS MWADAIKAFG LELNENADKE
    660 670 680 690 700
    RSTLSLQFVR GLGESPLARS YEASHASIAE NRELQSADSD RSTRHIEIAL
    710 720 730 740 750
    PPDVEYQEGD HLGVLPKNSQ TNVSRILHRF GLKGTDQVTL SASGRSAGHL
    760 770 780 790 800
    PLGRPVSLHD LLSYSVEVQE AATRAQIREL ASFTVCPPHR RELEELSAEG
    810 820 830 840 850
    VYQEQILKKR ISMLDLLEKY EACDMPFERF LELLRPLKPR YYSISSSPRV
    860 870 880 890 900
    NPRQASITVG VVRGPAWSGR GEYRGVASND LAERQAGDDV VMFIRTPESR
    910 920 930 940 950
    FQLPKDPETP IIMVGPGTGV APFRGFLQAR DVLKREGKTL GEAHLYFGCR
    960 970 980 990 1000
    NDRDFIYRDE LERFEKDGIV TVHTAFSRKE GMPKTYVQHL MADQADTLIS
    1010 1020 1030 1040 1050
    ILDRGGRLYV CGDGSKMAPD VEAALQKAYQ AVHGTGEQEA QNWLRHLQDT
    1060
    GMYAKDVWAG I
    Length:1,061
    Mass (Da):119,468
    Last modified:July 1, 1997 - v1
    Checksum:i7915DACC20578978
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D87979 Genomic DNA Translation: BAA20123.1
    AL009126 Genomic DNA Translation: CAB12544.1
    PIRiD69799
    RefSeqiNP_388606.1, NC_000964.3
    WP_003242884.1, NZ_JNCM01000032.1

    Genome annotation databases

    EnsemblBacteriaiCAB12544; CAB12544; BSU07250
    GeneIDi938784
    KEGGibsu:BSU07250
    PATRICifig|224308.179.peg.786

    Similar proteinsi

    Entry informationi

    Entry nameiCYPD_BACSU
    AccessioniPrimary (citable) accession number: O08394
    Entry historyiIntegrated into UniProtKB/Swiss-Prot: December 1, 2000
    Last sequence update: July 1, 1997
    Last modified: February 28, 2018
    This is version 141 of the entry and version 1 of the sequence. See complete history.
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    Complete proteome, Reference proteome
    UniProt is an ELIXIR core data resource
    Main funding by: National Institutes of Health