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Protein

Hydroxycinnamoyl-CoA hydratase-lyase

Gene
N/A
Organism
Pseudomonas fluorescens
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the hydration of the acyl-CoA thioester of ferulic acid and the subsequent retro-aldol cleavage of the hydrated intermediate to yield vanillin (4-hydroxy-3-methoxy-benzaldehyde).1 Publication

Catalytic activityi

4-hydroxy-3-methoxyphenyl-beta-hydroxypropanoyl-CoA = feruloyl-CoA + H2O.1 Publication
3-hydroxy-3-(4-hydroxy-3-methoxyphenyl)propanoyl-CoA = vanillin + acetyl-CoA.1 Publication

Kineticsi

kcat is 3.72 sec(-1) with ferulic acid as substrate.

  1. KM=0.0118 mM for ferulic acid1 Publication

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei29 – 291Coenzyme A
    Binding sitei75 – 751Substrate
    Binding sitei120 – 1201Coenzyme A; via carbonyl oxygen
    Binding sitei151 – 1511Substrate; via carbonyl oxygen
    Binding sitei239 – 2391Substrate

    GO - Molecular functioni

    Complete GO annotation...

    Keywords - Molecular functioni

    Lyase

    Enzyme and pathway databases

    BRENDAi4.2.1.101. 5121.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Hydroxycinnamoyl-CoA hydratase-lyase (EC:4.1.2.41, EC:4.2.1.101)
    Short name:
    HCHL
    Alternative name(s):
    P-hydroxycinnamoyl CoA hydratase/lyase
    Trans-feruloyl-CoA hydratase/vanillin synthase
    OrganismiPseudomonas fluorescens
    Taxonomic identifieri294 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaPseudomonadalesPseudomonadaceaePseudomonas

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi123 – 1231S → A: Reduced kcat compared to wild-type but not markerdly. 1 Publication
    Mutagenesisi143 – 1431E → A: Abolishes catalytic activity. 1 Publication
    Mutagenesisi239 – 2391Y → F: Increased KM but retains a significant amount of catalytic activity with a kcat 10 times less than that of the wild-type. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Initiator methionineiRemoved1 Publication
    Chaini2 – 276275Hydroxycinnamoyl-CoA hydratase-lyasePRO_0000418727Add
    BLAST

    Interactioni

    Subunit structurei

    Homohexamer; dimer of trimers.2 Publications

    Protein-protein interaction databases

    STRINGi1114970.PSF113_2659.

    Structurei

    Secondary structure

    1
    276
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi9 – 168Combined sources
    Beta strandi18 – 247Combined sources
    Helixi27 – 293Combined sources
    Helixi35 – 4915Combined sources
    Beta strandi54 – 629Combined sources
    Beta strandi65 – 673Combined sources
    Helixi72 – 8110Combined sources
    Helixi86 – 9813Combined sources
    Turni99 – 1046Combined sources
    Beta strandi109 – 1135Combined sources
    Beta strandi115 – 1184Combined sources
    Helixi119 – 1213Combined sources
    Helixi122 – 1276Combined sources
    Beta strandi128 – 1347Combined sources
    Beta strandi138 – 1403Combined sources
    Helixi142 – 1465Combined sources
    Helixi154 – 1618Combined sources
    Helixi164 – 17310Combined sources
    Beta strandi176 – 1783Combined sources
    Helixi179 – 1846Combined sources
    Beta strandi187 – 1926Combined sources
    Helixi194 – 20916Combined sources
    Helixi213 – 22614Combined sources
    Helixi231 – 24818Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    2J5IX-ray1.80A/B/C/D/E/F/G/H/I/J/K/L1-276[»]
    2VSSX-ray2.22A/B/C/D/E/F1-276[»]
    2VSUX-ray1.90A/B/C/D/E/F1-276[»]
    ProteinModelPortaliO69762.
    SMRiO69762. Positions 4-250.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiO69762.

    Family & Domainsi

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni70 – 723Coenzyme A binding
    Regioni142 – 1465Coenzyme A binding

    Sequence similaritiesi

    Phylogenomic databases

    eggNOGiENOG4108D7R. Bacteria.
    ENOG410XPS4. LUCA.

    Family and domain databases

    Gene3Di3.90.226.10. 1 hit.
    InterProiIPR029045. ClpP/crotonase-like_dom.
    IPR001753. Crotonase_core_superfam.
    IPR018376. Enoyl-CoA_hyd/isom_CS.
    [Graphical view]
    PfamiPF00378. ECH_1. 1 hit.
    [Graphical view]
    SUPFAMiSSF52096. SSF52096. 1 hit.
    PROSITEiPS00166. ENOYL_COA_HYDRATASE. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    O69762-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MSTYEGRWKT VKVEIEDGIA FVILNRPEKR NAMSPTLNRE MIDVLETLEQ
    60 70 80 90 100
    DPAAGVLVLT GAGEAWTAGM DLKEYFREVD AGPEILQEKI RREASQWQWK
    110 120 130 140 150
    LLRMYAKPTI AMVNGWCFGG GFSPLVACDL AICADEATFG LSEINWGIPP
    160 170 180 190 200
    GNLVSKAMAD TVGHRQSLYY IMTGKTFGGQ KAAEMGLVNE SVPLAQLREV
    210 220 230 240 250
    TIELARNLLE KNPVVLRAAK HGFKRCRELT WEQNEDYLYA KLDQSRLLDT
    260 270
    EGGREQGMKQ FLDDKSIKPG LQAYKR
    Length:276
    Mass (Da):31,008
    Last modified:August 1, 1998 - v1
    Checksum:iFDB5AD1539CA7F9A
    GO

    Experimental Info

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Sequence conflicti16 – 161E → Q AA sequence (PubMed:9461612).Curated

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    Y13067 Genomic DNA. Translation: CAA73502.1.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    Y13067 Genomic DNA. Translation: CAA73502.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    2J5IX-ray1.80A/B/C/D/E/F/G/H/I/J/K/L1-276[»]
    2VSSX-ray2.22A/B/C/D/E/F1-276[»]
    2VSUX-ray1.90A/B/C/D/E/F1-276[»]
    ProteinModelPortaliO69762.
    SMRiO69762. Positions 4-250.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi1114970.PSF113_2659.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Phylogenomic databases

    eggNOGiENOG4108D7R. Bacteria.
    ENOG410XPS4. LUCA.

    Enzyme and pathway databases

    BRENDAi4.2.1.101. 5121.

    Miscellaneous databases

    EvolutionaryTraceiO69762.

    Family and domain databases

    Gene3Di3.90.226.10. 1 hit.
    InterProiIPR029045. ClpP/crotonase-like_dom.
    IPR001753. Crotonase_core_superfam.
    IPR018376. Enoyl-CoA_hyd/isom_CS.
    [Graphical view]
    PfamiPF00378. ECH_1. 1 hit.
    [Graphical view]
    SUPFAMiSSF52096. SSF52096. 1 hit.
    PROSITEiPS00166. ENOYL_COA_HYDRATASE. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    1. "Metabolism of ferulic acid to vanillin. A bacterial gene of the enoyl-SCoA hydratase/isomerase superfamily encodes an enzyme for the hydration and cleavage of a hydroxycinnamic acid SCoA thioester."
      Gasson M.J., Kitamura Y., McLauchlan W.R., Narbad A., Parr A.J., Parsons E.L., Payne J., Rhodes M.J., Walton N.J.
      J. Biol. Chem. 273:4163-4170(1998) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-21, FUNCTION, CATALYTIC ACTIVITY.
      Strain: AN103.
    2. "The 1.8 A resolution structure of hydroxycinnamoyl-coenzyme A hydratase-lyase (HCHL) from Pseudomonas fluorescens, an enzyme that catalyses the transformation of feruloyl-coenzyme A to vanillin."
      Leonard P.M., Brzozowski A.M., Lebedev A., Marshall C.M., Smith D.J., Verma C.S., Walton N.J., Grogan G.
      Acta Crystallogr. D 62:1494-1501(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS), SUBUNIT.
      Strain: AN103.
    3. "A ternary complex of hydroxycinnamoyl-CoA hydratase-lyase (HCHL) with acetyl-CoA and vanillin gives insights into substrate specificity and mechanism."
      Bennett J.P., Bertin L., Moulton B., Fairlamb I.J., Brzozowski A.M., Walton N.J., Grogan G.
      Biochem. J. 414:281-289(2008) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH 4-HYDROXY-3-METHOXYBENZALDEHYDE AND ACETYL-COA, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, MUTAGENESIS OF SER-123; GLU-143 AND TYR-239.
      Strain: AN103.

    Entry informationi

    Entry nameiHCHL_PSEFL
    AccessioniPrimary (citable) accession number: O69762
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: September 5, 2012
    Last sequence update: August 1, 1998
    Last modified: November 11, 2015
    This is version 68 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    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

    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 one 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.