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Protein

2-alkenal reductase (NADP(+)-dependent)

Gene

DBR

Organism
Nicotiana tabacum (Common tobacco)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Reduces the C=C double bonds of alpha, beta unsaturated enones, but has no activity on enones with an endocyclic C=C double-bond. Shows a high specificity for NADPH as the hybrid donor. Substrates are 1-nitrocyclohexene, 2-methylpentenal, trans-cinnamaldehyde, methyl-trans-2-methylcinnamaldehyde, trans-2-nonenal and 1-octen-3-one. Reduced activity with aplha-methyl transcinnamaldehyde, 1-cyclohexene-1-carboxaldehyde, methyl crotonate, (R)-pulegone, and dimethyl itaconate and no activity with maleimides, citral, (5R)- or (5S)-carvone, (S)-perillyl alcohol, and substituted cyclohexenones and cyclopentenones (PubMed:17945329, Ref. 3). May also act as a allyl-alcohol dehydrogenase by catalyzing the dehydrogenation of secondary allylic alcohols rather than saturated secondary alcohols. Allyl-alcohol dehydrogenase is specific for the S-stereoisomer of the alcohols (PubMed:11117876).3 Publications

Catalytic activityi

An n-alkanal + NADP+ = an alk-2-enal + NADPH.
Allyl alcohol + NADP+ = acrolein + NADPH.

Kineticsi

kcat is 3.3 sec(-1) for (R)-pulegone at pH 7.0. kcat is 2.8 sec(-1) for (S)-pulegone at pH 7.0. kcat is 1.3 sec(-1) for NADPH at pH 7.3. kcat is 1.46 sec(-1) for 1-nitrocyclohexene at pH 7.3. kcat is 9.2 sec(-1) for 2-methylpentenal at pH 5.4. kcat is 0.66 sec(-1) for 2-methylpentenal at pH 7.3. kcat is 1.1 sec(-1) for methyl-trans-2-methylcinnamaldehyde at pH 7.3.

  1. KM=1.4 mM for (R)-pulegone (at pH 7.0)2 Publications
  2. KM=8.3 mM for (S)-pulegone (at pH 7.0)2 Publications
  3. KM=5.8 µM for NADPH (at pH 5.4)2 Publications
  4. KM=57 µM for 1-nitrocyclohexene (at pH 7.3)2 Publications
  5. KM=1032 µM for 2-methylpentenal (at pH 5.4)2 Publications
  6. KM=837 µM for 2-methylpentenal (at pH 7.3)2 Publications
  7. KM=516 µM for methyl-trans-2-methylcinnamaldehyde (at pH 7.3)2 Publications

    pH dependencei

    Optimum pH is 5.4 with 1-nitrocyclohexene as substrate.2 Publications

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei55 – 551SubstrateBy similarity
    Binding sitei80 – 801SubstrateBy similarity
    Binding sitei186 – 1861NADP; via amide nitrogen1 Publication
    Binding sitei190 – 1901NADP1 Publication
    Binding sitei206 – 2061NADP1 Publication
    Binding sitei230 – 2301NADP1 Publication
    Binding sitei252 – 2521NADP1 Publication
    Binding sitei258 – 2581NADP1 Publication
    Binding sitei332 – 3321NADP1 Publication

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi165 – 1662NADP1 Publication
    Nucleotide bindingi282 – 2843NADP1 Publication

    GO - Molecular functioni

    Complete GO annotation...

    Keywords - Molecular functioni

    Oxidoreductase

    Keywords - Ligandi

    NADP

    Enzyme and pathway databases

    BRENDAi1.1.1.54. 3645.
    1.3.1.102. 3645.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    2-alkenal reductase (NADP(+)-dependent) (EC:1.3.1.102)
    Alternative name(s):
    Alkenal double bound reductase
    Allyl-alcohol dehydrogenase (EC:1.1.1.54)
    Allylic alcohol dehydrogenase 1
    Short name:
    allyl-ADH1
    Flavin-free double bond reductase
    Short name:
    NtDBR
    Pulegone reductase
    Short name:
    NtRed-1
    Gene namesi
    Name:DBR
    OrganismiNicotiana tabacum (Common tobacco)
    Taxonomic identifieri4097 [NCBI]
    Taxonomic lineageiEukaryotaViridiplantaeStreptophytaEmbryophytaTracheophytaSpermatophytaMagnoliophytaeudicotyledonsGunneridaePentapetalaeasteridslamiidsSolanalesSolanaceaeNicotianoideaeNicotianeaeNicotiana

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 3433432-alkenal reductase (NADP(+)-dependent)PRO_0000423024Add
    BLAST

    Interactioni

    Subunit structurei

    Homodimer.3 Publications

    Structurei

    Secondary structure

    1
    343
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi4 – 129Combined sources
    Beta strandi17 – 193Combined sources
    Helixi22 – 243Combined sources
    Beta strandi25 – 339Combined sources
    Beta strandi43 – 519Combined sources
    Helixi54 – 607Combined sources
    Beta strandi78 – 9013Combined sources
    Beta strandi98 – 11114Combined sources
    Beta strandi118 – 1203Combined sources
    Helixi127 – 1315Combined sources
    Turni132 – 1343Combined sources
    Helixi136 – 14611Combined sources
    Turni147 – 1493Combined sources
    Beta strandi156 – 1616Combined sources
    Helixi167 – 17711Combined sources
    Beta strandi180 – 1878Combined sources
    Helixi188 – 1969Combined sources
    Beta strandi201 – 2055Combined sources
    Helixi206 – 2083Combined sources
    Helixi212 – 2198Combined sources
    Beta strandi224 – 2318Combined sources
    Helixi234 – 2407Combined sources
    Beta strandi243 – 2519Combined sources
    Helixi255 – 2573Combined sources
    Helixi271 – 2744Combined sources
    Beta strandi278 – 2814Combined sources
    Helixi284 – 2907Combined sources
    Helixi291 – 30313Combined sources
    Beta strandi311 – 3166Combined sources
    Helixi317 – 3193Combined sources
    Helixi320 – 3267Combined sources
    Helixi327 – 3293Combined sources
    Beta strandi333 – 3397Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    4HFJX-ray2.00A/B1-343[»]
    4HFMX-ray1.90A/B1-343[»]
    4HFNX-ray2.10A/B1-343[»]
    ProteinModelPortaliQ9SLN8.
    SMRiQ9SLN8. Positions 6-343.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Sequence similaritiesi

    Family and domain databases

    Gene3Di3.40.50.720. 1 hit.
    3.90.180.10. 1 hit.
    InterProiIPR013149. ADH_C.
    IPR002085. ADH_SF_Zn-type.
    IPR011032. GroES-like.
    IPR016040. NAD(P)-bd_dom.
    [Graphical view]
    PANTHERiPTHR11695. PTHR11695. 1 hit.
    PfamiPF00107. ADH_zinc_N. 1 hit.
    [Graphical view]
    SUPFAMiSSF50129. SSF50129. 2 hits.

    Sequencei

    Sequence statusi: Complete.

    Q9SLN8-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MAEEVSNKQV ILKNYVTGYP KESDMEIKNV TIKLKVPEGS NDVVVKNLYL
    60 70 80 90 100
    SCDPYMRSRM RKIEGSYVES FAPGSPITGY GVAKVLESGD PKFQKGDLVW
    110 120 130 140 150
    GMTGWEEYSI ITPTQTLFKI HDKDVPLSYY TGILGMPGMT AYAGFHEVCS
    160 170 180 190 200
    PKKGETVFVS AASGAVGQLV GQFAKMLGCY VVGSAGSKEK VDLLKSKFGF
    210 220 230 240 250
    DEAFNYKEEQ DLSAALKRYF PDGIDIYFEN VGGKMLDAVL VNMKLYGRIA
    260 270 280 290 300
    VCGMISQYNL EQTEGVHNLF CLITKRIRME GFLVFDYYHL YPKYLEMVIP
    310 320 330 340
    QIKAGKVVYV EDVAHGLESA PTALVGLFSG RNIGKQVVMV SRE
    Length:343
    Mass (Da):38,088
    Last modified:May 1, 2000 - v1
    Checksum:i235A567096F42637
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AB036735 mRNA. Translation: BAA89423.1.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    AB036735 mRNA. Translation: BAA89423.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    4HFJX-ray2.00A/B1-343[»]
    4HFMX-ray1.90A/B1-343[»]
    4HFNX-ray2.10A/B1-343[»]
    ProteinModelPortaliQ9SLN8.
    SMRiQ9SLN8. Positions 6-343.
    ModBaseiSearch...
    MobiDBiSearch...

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Enzyme and pathway databases

    BRENDAi1.1.1.54. 3645.
    1.3.1.102. 3645.

    Family and domain databases

    Gene3Di3.40.50.720. 1 hit.
    3.90.180.10. 1 hit.
    InterProiIPR013149. ADH_C.
    IPR002085. ADH_SF_Zn-type.
    IPR011032. GroES-like.
    IPR016040. NAD(P)-bd_dom.
    [Graphical view]
    PANTHERiPTHR11695. PTHR11695. 1 hit.
    PfamiPF00107. ADH_zinc_N. 1 hit.
    [Graphical view]
    SUPFAMiSSF50129. SSF50129. 2 hits.
    ProtoNetiSearch...

    Publicationsi

    1. "A 38 kDa allylic alcohol dehydrogenase from the cultured cells of Nicotiana tabacum."
      Hirata T., Tamura Y., Yokobatake N., Shimoda K., Ashida Y.
      Phytochemistry 55:297-303(2000) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 85-101 AND 153-178, FUNCTION, SUBUNIT.
    2. "An enone reductase from Nicotiana tabacum: cDNA cloning, expression in Escherichia coli, and reduction of enones with the recombinant proteins."
      Matsushima A., Sato Y., Otsuka M., Watanabe T., Yamamoto H., Hirata T.
      Bioorg. Chem. 36:23-28(2008) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 62-74, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT.
    3. "Stereospecific hydrogenation of the C C double bond of enones by Escherichia coli overexpressing an enone reductase of Nicotiana tabacum."
      Hirataa T., Matsushimab A., Satoa Y., Iwasakia T., Nomuraa H., Watanabea T., Toyodaa S., Izumia S.
      J. Mol. Catal., B Enzym. 59:158-162(2009)
      Cited for: FUNCTION, CATALYTIC ACTIVITY.
    4. "Finding Sequences for over 270 Orphan Enzymes."
      Shearer A.G., Altman T., Rhee C.D.
      PLoS ONE 9:E97250-E97250(2014) [PubMed] [Europe PMC] [Abstract]
      Cited for: IDENTIFICATION.
    5. "Biocatalytic asymmetric alkene reduction: crystal structure and characterization of a double bond reductase from Nicotiana tabacum."
      Mansell D.J., Toogood H.S., Waller J., Hughes J.M.X., Levy C.W., Gardiner J.M., Scrutton N.S.
      ACS Catal. 3:370-379(2013)
      Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) IN COMPLEX WITH NADPH AND SUBSTRATE, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT.

    Entry informationi

    Entry nameiDBR_TOBAC
    AccessioniPrimary (citable) accession number: Q9SLN8
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: July 24, 2013
    Last sequence update: May 1, 2000
    Last modified: April 1, 2015
    This is version 56 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programPlant Protein Annotation Program

    Miscellaneousi

    Miscellaneous

    There are no hydrogen bonds between the substrate tested and the crystalized protein (Ref. 5). The reduction of pulegone shows no sterospecificity and only the pro-4R hydrogen of NADPH is incorporated into the C=C double bond of pulegone (Ref. 3).

    Caution

    PubMed:11117876 showed an allyl-alcohol dehydrogenase activity on (2S,4S)-carveol while PubMed:17945329 and Ref. 5 showed an exclusive enone reductase activity.Curated

    Keywords - Technical termi

    3D-structure, Direct protein sequencing

    Documents

    1. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    2. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.