DBR_TOBAC
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 03-AUG-2022, entry version 89.
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 methyl-trans-
CC 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 iPTMnet; Q9SLN8; -.
DR GeneID; 107830910; -.
DR KEGG; ag:BAA89423; -.
DR KEGG; nta:107830910; -.
DR OMA; MADWPKN; -.
DR OrthoDB; 884151at2759; -.
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 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; PTHR43205; 1.
DR Pfam; PF16884; ADH_N_2; 1.
DR Pfam; PF00107; ADH_zinc_N; 1.
DR SMART; SM00829; PKS_ER; 1.
DR SUPFAM; SSF50129; SSF50129; 2.
DR SUPFAM; SSF51735; SSF51735; 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
