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Protein

Acetoacetyl-CoA reductase

Gene

phbB

Organism
Cupriavidus necator (strain ATCC 17699 / H16 / DSM 428 / Stanier 337) (Ralstonia eutropha)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the chiral reduction of acetoacetyl-CoA to (R)-3-hydroxybutyryl-CoA. Is involved in the biosynthesis of polyhydroxybutyrate (PHB), which is accumulated as an intracellular energy reserve material when cells grow under conditions of nutrient limitation.4 Publications

Catalytic activityi

(R)-3-hydroxyacyl-CoA + NADP+ = 3-oxoacyl-CoA + NADPH.
(R)-3-hydroxybutanoyl-CoA + NADP+ = acetoacetyl-CoA + NADPH.

Kineticsi

kcat is 102 sec(-1).

  1. KM=5.7 µM for acetoacetyl-CoA1 Publication
  2. KM=149 µM for NADPH1 Publication

    Pathwayi: poly-(R)-3-hydroxybutanoate biosynthesis

    This protein is involved in the pathway poly-(R)-3-hydroxybutanoate biosynthesis, which is part of Biopolymer metabolism.2 Publications
    View all proteins of this organism that are known to be involved in the pathway poly-(R)-3-hydroxybutanoate biosynthesis and in Biopolymer metabolism.

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei35 – 351NADP; via amide nitrogen1 Publication
    Binding sitei40 – 401NADP1 Publication
    Binding sitei94 – 941Substrate
    Active sitei153 – 1531Proton acceptorPROSITE-ProRule annotation
    Binding sitei195 – 1951Substrate

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi13 – 153NADP1 Publication
    Nucleotide bindingi60 – 623NADP1 Publication
    Nucleotide bindingi88 – 925NADP1 Publication
    Nucleotide bindingi183 – 1864NADP1 Publication

    GO - Molecular functioni

    GO - Biological processi

    Complete GO annotation...

    Keywords - Molecular functioni

    Oxidoreductase

    Keywords - Biological processi

    PHB biosynthesis

    Keywords - Ligandi

    NADP

    Enzyme and pathway databases

    BioCyciCNEC381666:GJUJ-1436-MONOMER.
    UniPathwayiUPA00917.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Acetoacetyl-CoA reductase (EC:1.1.1.36)
    Gene namesi
    Name:phbB
    Synonyms:phaB
    Ordered Locus Names:H16_A1439
    OrganismiCupriavidus necator (strain ATCC 17699 / H16 / DSM 428 / Stanier 337) (Ralstonia eutropha)
    Taxonomic identifieri381666 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaBetaproteobacteriaBurkholderialesBurkholderiaceaeCupriavidus
    Proteomesi
    • UP000008210 Componenti: Chromosome 1

    Subcellular locationi

    GO - Cellular componenti

    Complete GO annotation...

    Keywords - Cellular componenti

    Cytoplasm

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi47 – 471Q → L: 2.4-fold increase in activity. 2-fold decrease in affinity for NADPH and 2.8-fold decrease in affinity for acetoacetyl-CoA. 2 Publications
    Mutagenesisi94 – 941D → A: About 6% of wild-type activity. 1 Publication
    Mutagenesisi99 – 991K → A: Nearly loss of activity. 1 Publication
    Mutagenesisi147 – 1471Q → A: About 30% of wild-type activity. 1 Publication
    Mutagenesisi148 – 1481F → A: About 30% of wild-type activity. 1 Publication
    Mutagenesisi150 – 1501Q → A: About 20% of wild-type activity. 1 Publication
    Mutagenesisi173 – 1731T → S: 3.5-fold increase in activity. 4-fold decrease in affinity for NADPH and 2.4-fold decrease in affinity for acetoacetyl-CoA. 2 Publications
    Mutagenesisi185 – 1851Y → A: Nearly loss of activity. 1 Publication
    Mutagenesisi195 – 1951R → A: Nearly loss of activity. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 246246Acetoacetyl-CoA reductasePRO_0000054748Add
    BLAST

    Interactioni

    Subunit structurei

    Homotetramer.2 Publications

    Protein-protein interaction databases

    STRINGi381666.H16_A1439.

    Structurei

    Secondary structure

    1
    246
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi5 – 84Combined sources
    Turni9 – 124Combined sources
    Helixi14 – 2512Combined sources
    Beta strandi29 – 346Combined sources
    Helixi41 – 5010Combined sources
    Beta strandi56 – 594Combined sources
    Helixi65 – 7814Combined sources
    Beta strandi82 – 876Combined sources
    Helixi97 – 993Combined sources
    Helixi102 – 11211Combined sources
    Helixi114 – 13017Combined sources
    Beta strandi133 – 1386Combined sources
    Helixi141 – 1455Combined sources
    Helixi151 – 17121Combined sources
    Helixi172 – 1743Combined sources
    Beta strandi176 – 1838Combined sources
    Beta strandi185 – 1884Combined sources
    Helixi189 – 1924Combined sources
    Helixi196 – 2049Combined sources
    Helixi214 – 22512Combined sources
    Helixi227 – 2293Combined sources
    Beta strandi236 – 2405Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3VZPX-ray1.79A/B/C/D2-246[»]
    3VZQX-ray2.00A/B2-246[»]
    3VZRX-ray2.90A/B2-246[»]
    3VZSX-ray2.14A/B/C/D2-246[»]
    4N5LX-ray1.65A/B1-246[»]
    4N5MX-ray1.34A/B1-246[»]
    4N5NX-ray1.90A/B1-246[»]
    ProteinModelPortaliP14697.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni147 – 1504Substrate binding
    Regioni184 – 1852Substrate binding

    Sequence similaritiesi

    Phylogenomic databases

    eggNOGiENOG4105CHR. Bacteria.
    ENOG410XNW1. LUCA.
    KOiK00023.
    OMAiHLASENI.
    OrthoDBiEOG6N3CR8.

    Family and domain databases

    Gene3Di3.40.50.720. 1 hit.
    InterProiIPR011283. Acetoacetyl-CoA_reductase.
    IPR016040. NAD(P)-bd_dom.
    IPR020904. Sc_DH/Rdtase_CS.
    IPR002347. SDR_fam.
    [Graphical view]
    PANTHERiPTHR24322. PTHR24322. 2 hits.
    PfamiPF00106. adh_short. 1 hit.
    [Graphical view]
    PRINTSiPR00081. GDHRDH.
    PR00080. SDRFAMILY.
    SUPFAMiSSF51735. SSF51735. 1 hit.
    TIGRFAMsiTIGR01829. AcAcCoA_reduct. 1 hit.
    PROSITEiPS00061. ADH_SHORT. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    P14697-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MTQRIAYVTG GMGGIGTAIC QRLAKDGFRV VAGCGPNSPR REKWLEQQKA
    60 70 80 90 100
    LGFDFIASEG NVADWDSTKT AFDKVKSEVG EVDVLINNAG ITRDVVFRKM
    110 120 130 140 150
    TRADWDAVID TNLTSLFNVT KQVIDGMADR GWGRIVNISS VNGQKGQFGQ
    160 170 180 190 200
    TNYSTAKAGL HGFTMALAQE VATKGVTVNT VSPGYIATDM VKAIRQDVLD
    210 220 230 240
    KIVATIPVKR LGLPEEIASI CAWLSSEESG FSTGADFSLN GGLHMG
    Length:246
    Mass (Da):26,370
    Last modified:April 1, 1990 - v1
    Checksum:iAD6739E0F5C93C06
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    J04987 Genomic DNA. Translation: AAA21973.1.
    AM260479 Genomic DNA. Translation: CAJ92574.1.
    PIRiB34340. RDALAE.
    RefSeqiWP_010810131.1. NC_008313.1.

    Genome annotation databases

    EnsemblBacteriaiCAJ92574; CAJ92574; H16_A1439.
    GeneIDi4249784.
    KEGGireh:H16_A1439.
    PATRICi35232343. VBIRalEut6770_1828.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    J04987 Genomic DNA. Translation: AAA21973.1.
    AM260479 Genomic DNA. Translation: CAJ92574.1.
    PIRiB34340. RDALAE.
    RefSeqiWP_010810131.1. NC_008313.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3VZPX-ray1.79A/B/C/D2-246[»]
    3VZQX-ray2.00A/B2-246[»]
    3VZRX-ray2.90A/B2-246[»]
    3VZSX-ray2.14A/B/C/D2-246[»]
    4N5LX-ray1.65A/B1-246[»]
    4N5MX-ray1.34A/B1-246[»]
    4N5NX-ray1.90A/B1-246[»]
    ProteinModelPortaliP14697.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi381666.H16_A1439.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiCAJ92574; CAJ92574; H16_A1439.
    GeneIDi4249784.
    KEGGireh:H16_A1439.
    PATRICi35232343. VBIRalEut6770_1828.

    Phylogenomic databases

    eggNOGiENOG4105CHR. Bacteria.
    ENOG410XNW1. LUCA.
    KOiK00023.
    OMAiHLASENI.
    OrthoDBiEOG6N3CR8.

    Enzyme and pathway databases

    UniPathwayiUPA00917.
    BioCyciCNEC381666:GJUJ-1436-MONOMER.

    Family and domain databases

    Gene3Di3.40.50.720. 1 hit.
    InterProiIPR011283. Acetoacetyl-CoA_reductase.
    IPR016040. NAD(P)-bd_dom.
    IPR020904. Sc_DH/Rdtase_CS.
    IPR002347. SDR_fam.
    [Graphical view]
    PANTHERiPTHR24322. PTHR24322. 2 hits.
    PfamiPF00106. adh_short. 1 hit.
    [Graphical view]
    PRINTSiPR00081. GDHRDH.
    PR00080. SDRFAMILY.
    SUPFAMiSSF51735. SSF51735. 1 hit.
    TIGRFAMsiTIGR01829. AcAcCoA_reduct. 1 hit.
    PROSITEiPS00061. ADH_SHORT. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Poly-beta-hydroxybutyrate biosynthesis in Alcaligenes eutrophus H16. Characterization of the genes encoding beta-ketothiolase and acetoacetyl-CoA reductase."
      Peoples O.P., Sinskey A.J.
      J. Biol. Chem. 264:15293-15297(1989) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION, FUNCTION.
      Strain: ATCC 17699 / H16 / DSM 428 / Stanier 337.
    2. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: ATCC 17699 / H16 / DSM 428 / Stanier 337.
    3. "Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16. Identification and characterization of the PHB polymerase gene (phbC)."
      Peoples O.P., Sinskey A.J.
      J. Biol. Chem. 264:15298-15303(1989) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION IN PHB SYNTHESIS, PATHWAY.
      Strain: ATCC 17699 / H16 / DSM 428 / Stanier 337.
    4. "Directed evolution and structural analysis of NADPH-dependent acetoacetyl coenzyme A (acetoacetyl-CoA) reductase from Ralstonia eutropha reveals two mutations responsible for enhanced kinetics."
      Matsumoto K., Tanaka Y., Watanabe T., Motohashi R., Ikeda K., Tobitani K., Yao M., Tanaka I., Taguchi S.
      Appl. Environ. Microbiol. 79:6134-6139(2013) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.79 ANGSTROMS) OF WILD-TYPE AND MUTANTS LEU-47 AND SER-173 IN COMPLEX WITH ACETOACETYL-COENZYME A AND NADP, FUNCTION, CATALYTIC ACTIVITY, KINETIC PARAMETERS, SUBUNIT, PATHWAY, MUTAGENESIS OF GLN-47 AND THR-173.
      Strain: ATCC 17699 / H16 / DSM 428 / Stanier 337.
    5. "Crystal structure of (R)-3-hydroxybutyryl-CoA dehydrogenase PhaB from Ralstonia eutropha."
      Kim J., Chang J.H., Kim E.J., Kim K.J.
      Biochem. Biophys. Res. Commun. 443:783-788(2014) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.34 ANGSTROMS) OF APOENZYME AND IN COMPLEXES WITH ACETOACETYL-COENZYME A AND NADP, SUBUNIT, FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF GLN-47; ASP-94; LYS-99; GLN-147; PHE-148; GLN-150; THR-173; TYR-185 AND ARG-195.
      Strain: ATCC 17699 / H16 / DSM 428 / Stanier 337.

    Entry informationi

    Entry nameiPHBB_CUPNH
    AccessioniPrimary (citable) accession number: P14697
    Secondary accession number(s): Q0KBP7
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: April 1, 1990
    Last sequence update: April 1, 1990
    Last modified: April 13, 2016
    This is version 116 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, Complete proteome, Reference proteome

    Documents

    1. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    2. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    3. 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.