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Q9SLN8 (DBR_TOBAC) Reviewed, UniProtKB/Swiss-Prot

Last modified March 19, 2014. Version 52. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
2-alkenal reductase (NADP(+)-dependent)

EC=1.3.1.102
Alternative name(s):
Alkenal double bound reductase
Allylic alcohol dehydrogenase 1
Short name=allyl-ADH1
Flavin-free double bond reductase
Short name=NtDBR
Pulegone reductase
Short name=NtRed-1
Gene names
Name:DBR
OrganismNicotiana tabacum (Common tobacco)
Taxonomic identifier4097 [NCBI]
Taxonomic lineageEukaryotaViridiplantaeStreptophytaEmbryophytaTracheophytaSpermatophytaMagnoliophytaeudicotyledonsGunneridaePentapetalaeasteridslamiidsSolanalesSolanaceaeNicotianoideaeNicotianeaeNicotiana

Protein attributes

Sequence length343 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

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. Ref.2 Ref.3

Catalytic activity

An n-alkanal + NADP+ = an alk-2-enal + NADPH. Ref.2 Ref.3 Ref.4

Subunit structure

Homodimer. Ref.1 Ref.2 Ref.4

Miscellaneous

There are no hydrogen bonds between the substrate tested and the crystalized protein (Ref.4). 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).

Sequence similarities

Belongs to the NADP-dependent oxidoreductase L4BD family.

Caution

Ref.1 showed an allyl-alcohol dehydrogenase activity on (2S,4S)-carveol while Ref.2 and Ref.4 showed an exclusive enone reductase activity.

Biophysicochemical properties

Kinetic parameters:

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.

KM=1.4 mM for (R)-pulegone (at pH 7.0) Ref.2 Ref.4

KM=8.3 mM for (S)-pulegone (at pH 7.0)

KM=5.8 µM for NADPH (at pH 5.4)

KM=57 µM for 1-nitrocyclohexene (at pH 7.3)

KM=1032 µM for 2-methylpentenal (at pH 5.4)

KM=837 µM for 2-methylpentenal (at pH 7.3)

KM=516 µM for methyl-trans-2-methylcinnamaldehyde (at pH 7.3)

pH dependence:

Optimum pH is 5.4 with 1-nitrocyclohexene as substrate.

Ontologies

Keywords
   LigandNADP
   Molecular functionOxidoreductase
   Technical term3D-structure
Direct protein sequencing
Gene Ontology (GO)
   Molecular_functionoxidoreductase activity

Inferred from electronic annotation. Source: UniProtKB-KW

zinc ion binding

Inferred from electronic annotation. Source: InterPro

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 3433432-alkenal reductase (NADP(+)-dependent)
PRO_0000423024

Regions

Nucleotide binding165 – 1662NADP
Nucleotide binding282 – 2843NADP

Sites

Binding site551Substrate By similarity
Binding site801Substrate By similarity
Binding site1861NADP; via amide nitrogen
Binding site1901NADP
Binding site2061NADP
Binding site2301NADP
Binding site2521NADP
Binding site2581NADP
Binding site3321NADP

Secondary structure

.......................................................... 343
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
Q9SLN8 [UniParc].

Last modified May 1, 2000. Version 1.
Checksum: 235A567096F42637

FASTA34338,088
        10         20         30         40         50         60 
MAEEVSNKQV ILKNYVTGYP KESDMEIKNV TIKLKVPEGS NDVVVKNLYL SCDPYMRSRM 

        70         80         90        100        110        120 
RKIEGSYVES FAPGSPITGY GVAKVLESGD PKFQKGDLVW GMTGWEEYSI ITPTQTLFKI 

       130        140        150        160        170        180 
HDKDVPLSYY TGILGMPGMT AYAGFHEVCS PKKGETVFVS AASGAVGQLV GQFAKMLGCY 

       190        200        210        220        230        240 
VVGSAGSKEK VDLLKSKFGF DEAFNYKEEQ DLSAALKRYF PDGIDIYFEN VGGKMLDAVL 

       250        260        270        280        290        300 
VNMKLYGRIA VCGMISQYNL EQTEGVHNLF CLITKRIRME GFLVFDYYHL YPKYLEMVIP 

       310        320        330        340 
QIKAGKVVYV EDVAHGLESA PTALVGLFSG RNIGKQVVMV SRE 

« Hide

References

[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, 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]"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.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
AB036735 mRNA. Translation: BAA89423.1.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
4HFJX-ray2.00A/B1-343[»]
4HFMX-ray1.90A/B1-343[»]
4HFNX-ray2.10A/B1-343[»]
ProteinModelPortalQ9SLN8.
SMRQ9SLN8. Positions 6-343.
ModBaseSearch...
MobiDBSearch...

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Family and domain databases

Gene3D3.40.50.720. 1 hit.
3.90.180.10. 1 hit.
InterProIPR013149. ADH_C.
IPR002085. ADH_SF_Zn-type.
IPR011032. GroES-like.
IPR016040. NAD(P)-bd_dom.
[Graphical view]
PANTHERPTHR11695. PTHR11695. 1 hit.
PfamPF00107. ADH_zinc_N. 1 hit.
[Graphical view]
SUPFAMSSF50129. SSF50129. 2 hits.
ProtoNetSearch...

Entry information

Entry nameDBR_TOBAC
AccessionPrimary (citable) accession number: Q9SLN8
Entry history
Integrated into UniProtKB/Swiss-Prot: July 24, 2013
Last sequence update: May 1, 2000
Last modified: March 19, 2014
This is version 52 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programPlant Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references