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Protein

4-chlorobenzoyl coenzyme A dehalogenase

Gene
N/A
Organism
Pseudomonas sp. (strain CBS-3)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Dehalogenates 4-chlorobenzoyl-CoA, 4-iodobenzoyl-CoA and 4-bromobenzoyl-CoA, but not 4-fluorobenzoyl-CoA. Inactive towards crotonyl-CoA, alpha-methylcrotonyl-CoA and beta-methylcrotonyl-CoA.2 Publications

Catalytic activityi

4-chlorobenzoyl-CoA + H2O = 4-hydroxybenzoyl CoA + chloride.2 Publications

Enzyme regulationi

Inactivated by 1 mM Ag+ and by 5 mM Cu2+. Partially inhibited by 5 mM Zn2+, Mn2+, Co2+, Fe2+ and Ni2+. Unaffected by 10 mM Na+, K+ and Li+ and by 0.5 mM Mg2+, Mn2+, Fe2+, Ca2+, Co2+ and Zn2+. Inhibited by the sulfhydryl blocking agent 5,5'-dithio-bis-(2-nitrobenzoate), SDS and N-bromosuccinimide. Unaffected by sodium azide and EDTA. Inactivated following treatment with diethyl pyrocarbonate; this inactivation is reversible by treatment with hydroxylamine.3 Publications

Kineticsi

  1. KM=3.7 µM for 4-chlorobenzoyl-CoA8 Publications
  2. KM=4.2 µM for 4-bromobenzoyl-CoA8 Publications
  3. KM=6.5 µM for 4-iodobenzoyl-CoA8 Publications
  4. KM=10.4 µM for 2,4-dichlorobenzoyl-CoA8 Publications
  5. KM=42 µM for 3,4-dichlorobenzoyl-CoA8 Publications
  6. KM=30 µM for 4-chloro-2-nitrobenzoyl-CoA8 Publications
  7. KM=5.5 µM for 4-chloro-3-nitrobenzoyl-CoA8 Publications
  1. Vmax=2.2 µmol/min/mg enzyme with 4-chlorobenzoyl-CoA as substrate8 Publications

pH dependencei

Optimum pH is 10.0. Half maximum activity remains at pH 9.0 and 11.5. Stable from pH 6.5 to 11.0, activity is lost following 10 minutes incubation at pH 4.5 or 12.0.8 Publications

Temperature dependencei

Optimum temperature is 60 degrees Celsius. Retains 60% of maximum activity at 30 degrees Celsius and 65 degrees Celsius. After 15 minutes preincubation at 60 degrees Celsius 70% of the initial activity remains, 15 minutes preincubation at 65 degrees Celsius results in a total loss of activity.8 Publications

Pathwayi: 4-chlorobenzoate degradation

This protein is involved in step 2 of the subpathway that synthesizes 4-hydroxybenzoate from 4-chlorobenzoate.1 Publication
Proteins known to be involved in the 3 steps of the subpathway in this organism are:
  1. no protein annotated in this organism
  2. 4-chlorobenzoyl coenzyme A dehalogenase, 4-chlorobenzoate--CoA ligase
  3. 4-hydroxybenzoyl-CoA thioesterase
This subpathway is part of the pathway 4-chlorobenzoate degradation, which is itself part of Xenobiotic degradation.
View all proteins of this organism that are known to be involved in the subpathway that synthesizes 4-hydroxybenzoate from 4-chlorobenzoate, the pathway 4-chlorobenzoate degradation and in Xenobiotic degradation.

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei24 – 241Substrate1 Publication
Active sitei90 – 901Proton acceptor2 Publications
Binding sitei114 – 1141Substrate; via amide nitrogen
Active sitei145 – 1451Nucleophile2 Publications
Binding sitei257 – 2571Substrate; shared with oligomeric partner

GO - Molecular functioni

  • 4-chlorobenzoyl-CoA dehalogenase activity Source: UniProtKB

GO - Biological processi

  • coenzyme A metabolic process Source: UniProtKB
Complete GO annotation...

Keywords - Molecular functioni

Hydrolase

Enzyme and pathway databases

BioCyciMetaCyc:MONOMER-14753.
BRENDAi3.8.1.7. 5085.
SABIO-RKA5JTM5.
UniPathwayiUPA01011; UER01021.

Names & Taxonomyi

Protein namesi
Recommended name:
4-chlorobenzoyl coenzyme A dehalogenase1 Publication (EC:3.8.1.7)
Short name:
4-CBA-CoA dehalogenaseImported
Short name:
4-CBCoA dehalogenaseBy similarity
Short name:
4-chlorobenzoyl-CoA dehalogenase2 Publications
OrganismiPseudomonas sp. (strain CBS-3)
Taxonomic identifieri72586 [NCBI]
Taxonomic lineageiBacteriaProteobacteria

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi24 – 241R → K or L: Does not strongly affect catalytic activity, but reduces substrate CoA binding. 1 Publication
Mutagenesisi34 – 341E → T: Forms inclusion bodies. 1 Publication
Mutagenesisi43 – 431E → A: No effect on catalytic activity. 1 Publication
Mutagenesisi45 – 451D → A: No effect on catalytic activity. 1 Publication
Mutagenesisi46 – 461D → A: No effect on catalytic activity. 1 Publication
Mutagenesisi63 – 631G → A, I or P: Yields insoluble protein. 1 Publication
Mutagenesisi64 – 641F → A: 30-fold reduction in catalytic activity, substrate benzoyl group binding is unaffected. 3 Publications
Mutagenesisi64 – 641F → L: Retains catalytic activity, but substrate benzoyl group binding is decreased. 3 Publications
Mutagenesisi64 – 641F → P: Severely reduces catalytic activity. Arylated intermediate does not accumulate. 3 Publications
Mutagenesisi65 – 651Y → D: Catalytic activity is almost as efficient as wild type. 1 Publication
Mutagenesisi67 – 671R → K: Reduces substrate CoA binding. 1 Publication
Mutagenesisi67 – 671R → L: Forms inclusion bodies. 1 Publication
Mutagenesisi68 – 681E → T: No effect on catalytic activity. 1 Publication
Mutagenesisi81 – 811H → Q: Loss of catalytic activity, substrate benzoyl group binding is not affected. 2 Publications
Mutagenesisi82 – 821F → L: Retains catalytic activity, but substrate benzoyl group binding is decreased. 1 Publication
Mutagenesisi89 – 891W → F: Retains catalytic activity, but substrate benzoyl group binding is decreased. 1 Publication
Mutagenesisi89 – 891W → Y: Reduced activity and substrate benzoyl group binding. 1 Publication
Mutagenesisi90 – 901H → Q: Complete loss of catalytic activity (PubMed:8718880 and PubMed:9063883). Significantly reduced activity (PubMed:11695894). Substrate binding is not significantly affected. Reduced arylated intermediate formation. 3 Publications
Mutagenesisi94 – 941H → Q: No effect on catalytic activity. 1 Publication
Mutagenesisi112 – 1121A → G: Yields insoluble protein. 1 Publication
Mutagenesisi112 – 1121A → S: Protein precipitates upon purification. 1 Publication
Mutagenesisi112 – 1121A → V: Catalytic activity is almost as efficient as wild type. 1 Publication
Mutagenesisi113 – 1131G → A: Strongly reduced catalytic activity and substrate benzoyl group binding. Arylated intermediate does not accumulate. 2 Publications
Mutagenesisi113 – 1131G → N or S: Strongly reduced catalytic activity. Arylated intermediate does not accumulate. 2 Publications
Mutagenesisi113 – 1131G → V: Protein precipitates upon purification. 2 Publications
Mutagenesisi114 – 1141G → A: Strongly reduced catalytic activity and substrate benzoyl group binding. 2 Publications
Mutagenesisi114 – 1141G → P: Unstable. 2 Publications
Mutagenesisi115 – 1151G → L, N, S or V: Yields insoluble protein. 1 Publication
Mutagenesisi123 – 1231D → T: No effect on catalytic activity. 1 Publication
Mutagenesisi129 – 1291D → T: No effect on catalytic activity. 1 Publication
Mutagenesisi137 – 1371W → F: Low catalytic activity, but KM unaffected (PubMed:8718880). Retains catalytic activity, but substrate benzoyl group binding is decreased (PubMed:9063883). 2 Publications
Mutagenesisi145 – 1451D → A: Complete loss of catalytic activity, but not substrate binding. 1 Publication
Mutagenesisi163 – 1631E → T: No effect on catalytic activity. 1 Publication
Mutagenesisi175 – 1751E → D: No effect on catalytic activity. 1 Publication
Mutagenesisi179 – 1791W → F: No effect on catalytic activity. 1 Publication
Mutagenesisi208 – 2081H → Q: No effect on catalytic activity. 1 Publication
Mutagenesisi216 – 2161R → E, K or L: Yields insoluble protein. 1 Publication
Mutagenesisi232 – 2321E → A, N, Q or R: Yields insoluble protein. 1 Publication
Mutagenesisi232 – 2321E → D: Reduced catalytic activity, increased substrate binding. 1 Publication
Mutagenesisi257 – 2571R → K: Retains catalytic activity and substrate CoA binding. 1 Publication
Mutagenesisi257 – 2571R → L: Significantly reduces catalytic activity and substrate CoA binding. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 2692694-chlorobenzoyl coenzyme A dehalogenasePRO_0000401147Add
BLAST

Expressioni

Inductioni

By 4-chlorobenzoyl-CoA, 4-iodobenzoyl-CoA or 4-bromobenzoyl-CoA.1 Publication

Interactioni

Subunit structurei

Homotetramer (PubMed:1610806). Homotetramer, homooctamer and larger multimers (PubMed:7579994). Homotrimer (PubMed:8679561).4 Publications

Structurei

Secondary structure

1
269
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi3 – 108Combined sources
Beta strandi13 – 186Combined sources
Helixi21 – 233Combined sources
Helixi29 – 4416Combined sources
Beta strandi50 – 567Combined sources
Beta strandi59 – 613Combined sources
Helixi66 – 683Combined sources
Beta strandi71 – 733Combined sources
Helixi74 – 9825Combined sources
Beta strandi99 – 1013Combined sources
Beta strandi103 – 1075Combined sources
Beta strandi109 – 1124Combined sources
Helixi114 – 1218Combined sources
Beta strandi122 – 1287Combined sources
Beta strandi132 – 1343Combined sources
Helixi137 – 1404Combined sources
Helixi148 – 16720Combined sources
Helixi173 – 1786Combined sources
Beta strandi183 – 1864Combined sources
Helixi188 – 20417Combined sources
Helixi207 – 22014Combined sources
Helixi225 – 24117Combined sources
Helixi245 – 2539Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1JXZX-ray1.90A/B/C1-269[»]
1NZYX-ray1.80A/B/C1-269[»]
ProteinModelPortaliA5JTM5.
SMRiA5JTM5. Positions 1-269.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiA5JTM5.

Family & Domainsi

Region

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Regioni62 – 676Substrate binding

Sequence similaritiesi

Belongs to the enoyl-CoA hydratase/isomerase family.Sequence analysis

Phylogenomic databases

KOiK14418.

Family and domain databases

Gene3Di1.10.12.10. 1 hit.
3.90.226.10. 1 hit.
InterProiIPR029045. ClpP/crotonase-like_dom.
IPR014748. Crontonase_C.
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.

A5JTM5-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MYEAIGHRVE DGVAEITIKL PRHRNALSVK AMQEVTDALN RAEEDDSVGA
60 70 80 90 100
VMITGAEDAF CAGFYLREIP LDKGVAGVRD HFRIGALWWH QMIHKIIRVK
110 120 130 140 150
RPVLAAINGV AAGGGLGISL ASDMAICADS AKFVCAWHTI GIGNDTATSY
160 170 180 190 200
SLARIVGMRR AMELMLTNRT LYPEEAKDWG LVSRVYPKDD FREVAWKVAR
210 220 230 240 250
ELAAAPTHLQ VMAKERFHAG WMQPVEECTE FEIQNVIASV THPHFMPCLT
260
EFLDGHRADR PQVELPAGV
Length:269
Mass (Da):29,802
Last modified:June 26, 2007 - v1
Checksum:iEC165E9063C70B07
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
EF569604 Genomic DNA. Translation: ABQ44578.1.

Genome annotation databases

KEGGiag:ABQ44578.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
EF569604 Genomic DNA. Translation: ABQ44578.1.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1JXZX-ray1.90A/B/C1-269[»]
1NZYX-ray1.80A/B/C1-269[»]
ProteinModelPortaliA5JTM5.
SMRiA5JTM5. Positions 1-269.
ModBaseiSearch...
MobiDBiSearch...

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Genome annotation databases

KEGGiag:ABQ44578.

Phylogenomic databases

KOiK14418.

Enzyme and pathway databases

UniPathwayiUPA01011; UER01021.
BioCyciMetaCyc:MONOMER-14753.
BRENDAi3.8.1.7. 5085.
SABIO-RKA5JTM5.

Miscellaneous databases

EvolutionaryTraceiA5JTM5.

Family and domain databases

Gene3Di1.10.12.10. 1 hit.
3.90.226.10. 1 hit.
InterProiIPR029045. ClpP/crotonase-like_dom.
IPR014748. Crontonase_C.
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. "Ancestry of the 4-chlorobenzoate dehalogenase: analysis of amino acid sequence identities among families of acyl:adenyl ligases, enoyl-CoA hydratases/isomerases, and acyl-CoA thioesterases."
    Babbitt P.C., Kenyon G.L., Martin B.M., Charest H., Slyvestre M., Scholten J.D., Chang K.H., Liang P.H., Dunaway-Mariano D.
    Biochemistry 31:5594-5604(1992) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
  2. "The nucleotide sequence upstream and downstream of 4-CBA-CoA ORFs in 9.5 kb Pseudomonas sp. strain CBS 3 chromosomal DNA."
    Zhang W., Dunaway-Mariano D.
    Submitted (APR-2007) to the EMBL/GenBank/DDBJ databases
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    Strain: CBS-3.
  3. "Dehalogenation of 4-chlorobenzoate. Characterisation of 4-chlorobenzoyl-coenzyme A dehalogenase from Pseudomonas sp. CBS3."
    Loffler F., Lingens F., Muller R.
    Biodegradation 6:203-212(1995) [PubMed] [Europe PMC] [Abstract]
    Cited for: PROTEIN SEQUENCE OF 1-40, FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, INDUCTION.
    Strain: CBS-3.
  4. "Isolation and characterization of the three polypeptide components of 4-chlorobenzoate dehalogenase from Pseudomonas sp. strain CBS-3."
    Chang K.H., Liang P.H., Beck W., Scholten J.D., Dunaway-Mariano D.
    Biochemistry 31:5605-5610(1992) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY, SUBUNIT.
    Strain: CBS-3.
  5. "Specificity of 4-chlorobenzoyl coenzyme A dehalogenase catalyzed dehalogenation of halogenated aromatics."
    Liang P.H., Yang G., Dunaway-Mariano D.
    Biochemistry 32:12245-12250(1993) [PubMed] [Europe PMC] [Abstract]
    Cited for: BIOPHYSICOCHEMICAL PROPERTIES.
  6. "Identification of active site residues essential to 4-chlorobenzoyl-coenzyme A dehalogenase catalysis by chemical modification and site directed mutagenesis."
    Yang G., Liu R.Q., Taylor K.L., Xiang H., Price J., Dunaway-Mariano D.
    Biochemistry 35:10879-10885(1996) [PubMed] [Europe PMC] [Abstract]
    Cited for: ENZYME REGULATION, ACTIVE SITES, MUTAGENESIS OF GLU-34; GLU-43; ASP-45; ASP-46; GLU-68; HIS-90; HIS-94; ASP-123; ASP-129; TRP-137; GLU-163; GLU-175; TRP-179 AND HIS-208.
  7. "Investigation of substrate activation by 4-chlorobenzoyl-coenzyme A dehalogenase."
    Taylor K.L., Xiang H., Liu R.Q., Yang G., Dunaway-Mariano D.
    Biochemistry 36:1349-1361(1997) [PubMed] [Europe PMC] [Abstract]
    Cited for: BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF PHE-64; HIS-81; PHE-82; TRP-89; HIS-90; GLY-114; TRP-137 AND ASP-145.
  8. "Role of active site binding interactions in 4-chlorobenzoyl-coenzyme A dehalogenase catalysis."
    Luo L., Taylor K.L., Xiang H., Wei Y., Zhang W., Dunaway-Mariano D.
    Biochemistry 40:15684-15692(2001) [PubMed] [Europe PMC] [Abstract]
    Cited for: BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ARG-24; PHE-64; ARG-67; GLY-113; GLY-114 AND ARG-257.
  9. "The strength of dehalogenase-substrate hydrogen bonding correlates with the rate of Meisenheimer intermediate formation."
    Dong J., Lu X., Wei Y., Luo L., Dunaway-Mariano D., Carey P.R.
    Biochemistry 42:9482-9490(2003) [PubMed] [Europe PMC] [Abstract]
    Cited for: BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF GLY-63; PHE-64; TYR-65; ALA-112; GLY-113 AND GLY-115.
  10. "Contributions of long-range electrostatic interactions to 4-chlorobenzoyl-CoA dehalogenase catalysis: a combined theoretical and experimental study."
    Wu J., Xu D., Lu X., Wang C., Guo H., Dunaway-Mariano D.
    Biochemistry 45:102-112(2006) [PubMed] [Europe PMC] [Abstract]
    Cited for: BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ARG-216 AND GLU-232.
  11. "Structure of 4-chlorobenzoyl coenzyme A dehalogenase determined to 1.8 A resolution: an enzyme catalyst generated via adaptive mutation."
    Benning M.M., Taylor K.L., Liu R.-Q., Yang G., Xiang H., Wesenberg G., Dunaway-Mariano D., Holden H.M.
    Biochemistry 35:8103-8109(1996) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF MUTANT GLN-90 IN COMPLEX WITH 4-HYDROXYBENZOYL COENZYME A, SUBUNIT, ACTIVE SITE, SUBSTRATE-BINDING SITES.
    Strain: CBS-3.
  12. "Histidine 90 function in 4-chlorobenzoyl-coenzyme a dehalogenase catalysis."
    Zhang W., Wei Y., Luo L., Taylor K.L., Yang G., Dunaway-Mariano D., Benning M.M., Holden H.M.
    Biochemistry 40:13474-13482(2001) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF MUTANT GLN-90 IN COMPLEX WITH 4-HYDROXYBENZOYL COENZYME A, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF HIS-90.

Entry informationi

Entry nameiCBADH_PSEUC
AccessioniPrimary (citable) accession number: A5JTM5
Entry historyi
Integrated into UniProtKB/Swiss-Prot: November 30, 2010
Last sequence update: June 26, 2007
Last modified: November 11, 2015
This is version 42 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. 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.