ID   DBR_TOBAC               Reviewed;         343 AA.
AC   Q9SLN8;
DT   24-JUL-2013, integrated into UniProtKB/Swiss-Prot.
DT   01-MAY-2000, sequence version 1.
DT   27-MAR-2024, entry version 95.
DE   RecName: Full=2-alkenal reductase (NADP(+)-dependent);
DE            EC=1.3.1.102 {ECO:0000269|PubMed:17945329, ECO:0000269|Ref.3, ECO:0000269|Ref.4};
DE   AltName: Full=Alkenal double bound reductase;
DE   AltName: Full=Allylic alcohol dehydrogenase 1 {ECO:0000303|PubMed:11117876};
DE            Short=allyl-ADH1 {ECO:0000303|PubMed:11117876};
DE   AltName: Full=Flavin-free double bond reductase {ECO:0000303|Ref.4};
DE            Short=NtDBR;
DE   AltName: Full=Pulegone reductase {ECO:0000303|Ref.3};
DE            Short=NtRed-1;
GN   Name=DBR;
OS   Nicotiana tabacum (Common tobacco).
OC   Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC   Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC   asterids; lamiids; Solanales; Solanaceae; Nicotianoideae; Nicotianeae;
OC   Nicotiana.
OX   NCBI_TaxID=4097;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 85-101 AND 153-178,
RP   FUNCTION, AND SUBUNIT.
RX   PubMed=11117876; DOI=10.1016/s0031-9422(00)00326-5;
RA   Hirata T., Tamura Y., Yokobatake N., Shimoda K., Ashida Y.;
RT   "A 38 kDa allylic alcohol dehydrogenase from the cultured cells of
RT   Nicotiana tabacum.";
RL   Phytochemistry 55:297-303(2000).
RN   [2]
RP   PROTEIN SEQUENCE OF 62-74, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP   PROPERTIES, AND SUBUNIT.
RX   PubMed=17945329; DOI=10.1016/j.bioorg.2007.08.005;
RA   Matsushima A., Sato Y., Otsuka M., Watanabe T., Yamamoto H., Hirata T.;
RT   "An enone reductase from Nicotiana tabacum: cDNA cloning, expression in
RT   Escherichia coli, and reduction of enones with the recombinant proteins.";
RL   Bioorg. Chem. 36:23-28(2008).
RN   [3]
RP   FUNCTION, AND CATALYTIC ACTIVITY.
RX   DOI=10.1016/j.molcatb.2009.02.009;
RA   Hirataa T., Matsushimab A., Satoa Y., Iwasakia T., Nomuraa H.,
RA   Watanabea T., Toyodaa S., Izumia S.;
RT   "Stereospecific hydrogenation of the C=C double bond of enones by
RT   Escherichia coli overexpressing an enone reductase of Nicotiana tabacum.";
RL   J. Mol. Catal., B Enzym. 59:158-162(2009).
RN   [4]
RP   X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) IN COMPLEX WITH NADPH AND SUBSTRATE,
RP   CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX   DOI=10.1021/cs300709m;
RA   Mansell D.J., Toogood H.S., Waller J., Hughes J.M.X., Levy C.W.,
RA   Gardiner J.M., Scrutton N.S.;
RT   "Biocatalytic asymmetric alkene reduction: crystal structure and
RT   characterization of a double bond reductase from Nicotiana tabacum.";
RL   ACS Catal. 3:370-379(2013).
CC   -!- FUNCTION: Reduces the C=C double bonds of alpha, beta unsaturated
CC       enones, but has no activity on enones with an endocyclic C=C double-
CC       bond. Shows a high specificity for NADPH as the hybrid donor.
CC       Substrates are 1-nitrocyclohexene, 2-methylpentenal, trans-
CC       cinnamaldehyde, methyl-trans-2-methylcinnamaldehyde, trans-2-nonenal
CC       and 1-octen-3-one. Reduced activity with aplha-methyl
CC       transcinnamaldehyde, 1-cyclohexene-1-carboxaldehyde, methyl crotonate,
CC       (R)-pulegone, and dimethyl itaconate and no activity with maleimides,
CC       citral, (5R)- or (5S)-carvone, (S)-perillyl alcohol, and substituted
CC       cyclohexenones and cyclopentenones (PubMed:17945329, Ref.3). May also
CC       act as a allyl-alcohol dehydrogenase by catalyzing the dehydrogenation
CC       of secondary allylic alcohols rather than saturated secondary alcohols.
CC       Allyl-alcohol dehydrogenase is specific for the S-stereoisomer of the
CC       alcohols (PubMed:11117876). {ECO:0000269|PubMed:11117876,
CC       ECO:0000269|PubMed:17945329, ECO:0000269|Ref.3}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=an n-alkanal + NADP(+) = an alk-2-enal + H(+) + NADPH;
CC         Xref=Rhea:RHEA:13737, ChEBI:CHEBI:12834, ChEBI:CHEBI:13757,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC         EC=1.3.1.102; Evidence={ECO:0000269|PubMed:17945329,
CC         ECO:0000269|Ref.3, ECO:0000269|Ref.4};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=1.4 mM for (R)-pulegone (at pH 7.0) {ECO:0000269|PubMed:17945329,
CC         ECO:0000269|Ref.4};
CC         KM=8.3 mM for (S)-pulegone (at pH 7.0) {ECO:0000269|PubMed:17945329,
CC         ECO:0000269|Ref.4};
CC         KM=5.8 uM for NADPH (at pH 5.4) {ECO:0000269|PubMed:17945329,
CC         ECO:0000269|Ref.4};
CC         KM=57 uM for 1-nitrocyclohexene (at pH 7.3)
CC         {ECO:0000269|PubMed:17945329, ECO:0000269|Ref.4};
CC         KM=1032 uM for 2-methylpentenal (at pH 5.4)
CC         {ECO:0000269|PubMed:17945329, ECO:0000269|Ref.4};
CC         KM=837 uM for 2-methylpentenal (at pH 7.3)
CC         {ECO:0000269|PubMed:17945329, ECO:0000269|Ref.4};
CC         KM=516 uM for methyl-trans-2-methylcinnamaldehyde (at pH 7.3)
CC         {ECO:0000269|PubMed:17945329, ECO:0000269|Ref.4};
CC         Note=kcat is 3.3 sec(-1) for (R)-pulegone at pH 7.0. kcat is 2.8
CC         sec(-1) for (S)-pulegone at pH 7.0. kcat is 1.3 sec(-1) for NADPH at
CC         pH 7.3. kcat is 1.46 sec(-1) for 1-nitrocyclohexene at pH 7.3. kcat
CC         is 9.2 sec(-1) for 2-methylpentenal at pH 5.4. kcat is 0.66 sec(-1)
CC         for 2-methylpentenal at pH 7.3. kcat is 1.1 sec(-1) for
CC         methyl-trans-2-methylcinnamaldehyde at pH 7.3.;
CC       pH dependence:
CC         Optimum pH is 5.4 with 1-nitrocyclohexene as substrate.
CC         {ECO:0000269|PubMed:17945329, ECO:0000269|Ref.4};
CC   -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11117876,
CC       ECO:0000269|PubMed:17945329, ECO:0000269|Ref.4}.
CC   -!- MISCELLANEOUS: There are no hydrogen bonds between the substrate tested
CC       and the crystalized protein (Ref.4). The reduction of pulegone shows no
CC       sterospecificity and only the pro-4R hydrogen of NADPH is incorporated
CC       into the C=C double bond of pulegone (Ref.3).
CC   -!- SIMILARITY: Belongs to the NADP-dependent oxidoreductase L4BD family.
CC       {ECO:0000305}.
CC   -!- CAUTION: PubMed:11117876 showed an allyl-alcohol dehydrogenase activity
CC       on (2S,4S)-carveol while PubMed:17945329 and Ref.4 showed an exclusive
CC       enone reductase activity. {ECO:0000305}.
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DR   EMBL; AB036735; BAA89423.1; -; mRNA.
DR   RefSeq; NP_001313179.1; NM_001326250.1.
DR   RefSeq; XP_016514079.1; XM_016658593.1.
DR   PDB; 4HFJ; X-ray; 2.00 A; A/B=1-343.
DR   PDB; 4HFM; X-ray; 1.90 A; A/B=1-343.
DR   PDB; 4HFN; X-ray; 2.10 A; A/B=1-343.
DR   PDBsum; 4HFJ; -.
DR   PDBsum; 4HFM; -.
DR   PDBsum; 4HFN; -.
DR   AlphaFoldDB; Q9SLN8; -.
DR   SMR; Q9SLN8; -.
DR   STRING; 4097.Q9SLN8; -.
DR   iPTMnet; Q9SLN8; -.
DR   PaxDb; 4097-Q9SLN8; -.
DR   GeneID; 107830910; -.
DR   KEGG; ag:BAA89423; -.
DR   KEGG; nta:107830910; -.
DR   OMA; YIRDGTI; -.
DR   OrthoDB; 179761at2759; -.
DR   BRENDA; 1.1.1.54; 3645.
DR   BRENDA; 1.3.1.102; 3645.
DR   Proteomes; UP000084051; Unplaced.
DR   GO; GO:0032440; F:2-alkenal reductase [NAD(P)+] activity; IBA:GO_Central.
DR   GO; GO:0008152; P:metabolic process; IEA:UniProt.
DR   CDD; cd08295; double_bond_reductase_like; 1.
DR   Gene3D; 3.90.180.10; Medium-chain alcohol dehydrogenases, catalytic domain; 1.
DR   Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1.
DR   InterPro; IPR013149; ADH-like_C.
DR   InterPro; IPR041694; ADH_N_2.
DR   InterPro; IPR011032; GroES-like_sf.
DR   InterPro; IPR045010; MDR_fam.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   InterPro; IPR020843; PKS_ER.
DR   PANTHER; PTHR43205:SF86; ALLYL ALCOHOL DEHYDROGENASE_ NADP-DEPENDENT OXIDOREDUCTASE-LIKE PROTEIN-RELATED; 1.
DR   PANTHER; PTHR43205; PROSTAGLANDIN REDUCTASE; 1.
DR   Pfam; PF16884; ADH_N_2; 1.
DR   Pfam; PF00107; ADH_zinc_N; 1.
DR   SMART; SM00829; PKS_ER; 1.
DR   SUPFAM; SSF50129; GroES-like; 2.
DR   SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Direct protein sequencing; NADP; Oxidoreductase;
KW   Reference proteome.
FT   CHAIN           1..343
FT                   /note="2-alkenal reductase (NADP(+)-dependent)"
FT                   /id="PRO_0000423024"
FT   BINDING         55
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         80
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250"
FT   BINDING         165..166
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         186
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         190
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         206
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         230
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         252
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         258
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         282..284
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   BINDING         332
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000269|Ref.4"
FT   STRAND          4..12
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          17..19
FT                   /evidence="ECO:0007829|PDB:4HFJ"
FT   HELIX           22..24
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          25..33
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          43..51
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           54..60
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          78..90
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          98..111
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          118..120
FT                   /evidence="ECO:0007829|PDB:4HFJ"
FT   HELIX           127..131
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   TURN            132..134
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           136..146
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   TURN            147..149
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          156..161
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           167..177
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          180..187
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           188..196
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          201..205
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           206..208
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           212..219
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          224..231
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           234..240
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          243..251
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           255..257
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           271..274
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          278..281
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           284..290
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           291..303
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          311..316
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           317..319
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           320..326
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   HELIX           327..329
FT                   /evidence="ECO:0007829|PDB:4HFM"
FT   STRAND          333..339
FT                   /evidence="ECO:0007829|PDB:4HFM"
SQ   SEQUENCE   343 AA;  38088 MW;  235A567096F42637 CRC64;
     MAEEVSNKQV ILKNYVTGYP KESDMEIKNV TIKLKVPEGS NDVVVKNLYL SCDPYMRSRM
     RKIEGSYVES FAPGSPITGY GVAKVLESGD PKFQKGDLVW GMTGWEEYSI ITPTQTLFKI
     HDKDVPLSYY TGILGMPGMT AYAGFHEVCS PKKGETVFVS AASGAVGQLV GQFAKMLGCY
     VVGSAGSKEK VDLLKSKFGF DEAFNYKEEQ DLSAALKRYF PDGIDIYFEN VGGKMLDAVL
     VNMKLYGRIA VCGMISQYNL EQTEGVHNLF CLITKRIRME GFLVFDYYHL YPKYLEMVIP
     QIKAGKVVYV EDVAHGLESA PTALVGLFSG RNIGKQVVMV SRE
//