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Information on EC 4.2.1.17 - enoyl-CoA hydratase and Organism(s) Rattus norvegicus
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4.2.1.17 enoyl-CoA hydrataseIUBMB Comments
Acts in the reverse direction. With cis-compounds, yields (3R)-3-hydroxyacyl-CoA. cf. EC 4.2.1.74 long-chain-enoyl-CoA hydratase.
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Word Map
- 4.2.1.17
-
beta-oxidation
-
thiolase
- 3-ketoacyl-coa
-
trifunctional
- l-3-hydroxyacyl-coa
- carnitine
- crotonase
- leigh
- palmitoyl-coa
-
2,4-dienoyl-coa
- clofibrate
- polyhydroxyalkanoate
-
2-trans-enoyl-coas
- acetoacetyl-coa
- proliferators
-
r-specific
-
1.1.1.35
-
leigh-like
- 3-oxoacyl-coa
-
d-bifunctional
- octanoyl-coa
-
hydratase/3-hydroxyacyl-coa
- medicine
-
even-numbered
The taxonomic range for the selected organisms is: Rattus norvegicus
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
enoyl-coa hydratase, echs1, peroxisomal bifunctional enzyme, 2-enoyl-coa hydratase, amech, short-chain enoyl-coa hydratase, enoyl-coa hydratase/isomerase, beta-hydroxyacyl-coa dehydrase, enoyl-coenzyme a hydratase, fadb', 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
2-enoyl-CoA hydratase
-
-
-
-
2-enoyl-CoA hydratase 1
2-octenoyl coenzyme A hydrase
-
-
-
-
acyl coenzyme A hydrase
-
-
-
-
beta-hydroxyacid dehydrase
-
-
-
-
beta-hydroxyacyl-CoA dehydrase
-
-
-
-
crotonase
crotonyl hydrase
-
-
-
-
D-3-hydroxyacyl-CoA dehydratase
-
-
-
-
ECH
enol-CoA hydratase
-
-
-
-
Enoyl coenzyme A hydrase (D)
-
-
-
-
enoyl coenzyme A hydrase (L)
-
-
-
-
enoyl coenzyme A hydratase
-
-
-
-
enoyl hydrase
-
-
-
-
enoyl-CoA hydratase
enoyl-CoA hydratase 1
hydratase, enoyl coenzyme A
-
-
-
-
mitochondrial enoyl coenzyme A hydratase
the classification is ambiguous because the stereochemistry is not exactly determined
perMFE-1
perMFE-I
-
peroxisomal multifunctional enzyme perMFE-I has 2-enoyl-CoA hydratase 1 activity (L-specific, EC 4.2.1.17) and L-specific 3-hydroxyacyl-CoA dehydrogenase (1.1.1.35) activity. Peroxisomal multifunctional enzyme perMFE-II has 2-enoyl-CoA hydratase 2 (D-specific) activity and D-specific 3-hydroxyacyl-CoA dehydrogenase (1.1.1.36) activity
SCEH
short chain enoyl coenzyme A hydratase
-
-
-
-
short-chain enoyl-CoA hydratase
-
-
-
-
trans-2-enoyl-CoA hydratase
-
-
-
-
unsaturated acyl-CoA hydratase
-
-
-
-
2-enoyl-CoA hydratase 1
-
is part of peroxisomal multifunctional enzyme perMFE-I together with L-specific 3-hydroxyacyl-CoA dehydrogenase (1.1.1.35)
crotonase
-
-
-
-
ECH
-
-
-
-
enoyl-CoA hydratase
-
the classification is ambiguous because the stereochemistry is not exactly determined
enoyl-CoA hydratase
the classification is ambiguous because the stereochemistry is not exactly determined
perMFE-1
-
multifunctional enzyme, cf. 5.3.3.8 and EC 1.1.1.35, second multifunctional enzyme in rat liver peroxisome perMFE-2, cf. EC 4.2.1.107 and EC 4.2.1.119
SCEH
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(3S)-3-hydroxyacyl-CoA = trans-2(or 3)-enoyl-CoA + H2O
the CoA moiety of the substrate adopts the typical horseshoe conformation in a binding pocket formed from two adjacent monomers, while the active site catalytic residues are provided by a single monomer. The active site of rat liver mitochondrial ECH contains an oxyanion hole formed from the backbone amides of Gly141 and Ala98, which serves to polarize the carbonyl group of the alpha,beta-unsaturated enoyl-CoA substrate and stabilize the enolate intermediate. Two active site glutamate residues (Glu144 and Glu164) are proposed to act as a general base(s) for the conjugate addition of water onto the substrate enone and to protonate the resultant enolate
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
elimination
-
-
-
-
hydration
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
alpha-Linolenic acid metabolism, Aminobenzoate degradation, Benzoate degradation, beta-Alanine metabolism, Biosynthesis of secondary metabolites, Butanoate metabolism, Caprolactam degradation, Carbon fixation pathways in prokaryotes, Fatty acid degradation, Fatty acid elongation, Geraniol degradation, Limonene and pinene degradation, Lysine degradation, Microbial metabolism in diverse environments, Phenylalanine metabolism, Propanoate metabolism, Tryptophan metabolism, Valine, leucine and isoleucine degradation
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-, -, -, -, -, -, -, -, -, -, -, -, -, -, -, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
(3S)-3-hydroxyacyl-CoA hydro-lyase
Acts in the reverse direction. With cis-compounds, yields (3R)-3-hydroxyacyl-CoA. cf. EC 4.2.1.74 long-chain-enoyl-CoA hydratase.
CAS REGISTRY NUMBER
COMMENTARY
9027-13-8
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(Z)-2-butenoyl-CoA + H2O
(3R)-3-hydroxybutanoyl-CoA
-
kcat is 12fold slower than with the trans-iosmer crotonyl-CoA
-
-
?
3-octynoyl-CoA + H2O
3-ketooctanoyl-CoA
-
reaction of ECH1, ECH2 is inactivated by the compound, it is possible that 3-octynoyl-CoA is isomerized to reactive 2,3-octadienoyl-CoA, overview
-
-
?
methacrylyl-CoA + H2O
3-hydroxy-2-methylpropanoyl-CoA
-
-
-
r
trans-2-hexadecenoyl-CoA + H2O
(3S)-3-hydroxyhexadecanoyl-CoA + (3R)-3-hydroxyhexadecanoyl-CoA
-
rat liver homogenate enzyme activity is (S)-specific
(3S)-3-hydroxyhexadecanoyl-CoA is the dominant product
-
?
(2E)-enoyl-CoA + H2O
(3S)-hydroxyacyl-CoA
2E-enoyl-CoA is the product of the DELTA3,DELTA2-enoyl-CoA isomerase (EC 5.3.3.8) reaction, which subsequently is converted into (3S)-hydroxyacyl-CoA in the hydration step
-
-
?
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
-
i.e. (E)-2-butenoyl-CoA. The reaction proceeds via the syn addition of water and thus the pro-2R proton of (3S)-hydroxybutyryl-CoA is derived from solvent. The equilibrium constant for the hydration of trans-2-crotonyl-CoA to (3S)-hydroxybutyryl-CoA is 7.5. The rate of 3(R)-hydroxybutyryl-CoA formation is 400000fold slower than the normal hydration reaction (of crotonyl-CoA to (3S)-3-hydroxybutanoyl-CoA) but at least 1600000fold faster than the non-enzyme-catalyzed reaction. Formation of the incorrect stereoisomer likely occurs via syn addition of water to the incorrect face of the trans-2-crotonyl-CoA double bond. The absolute stereospecificity for the enzyme-catalyzed reaction is 1 in 400000. To account for the exchange of the hydroxybutyryl pro-2S proton, the enzyme must also catalyze the dehydration of 3(R)-hydroxybutyryl-CoA to cis-2-crotonyl-CoA. Thus, the enzyme is capable of catalyzing the epimerization of hydroxybutyryl-CoA
-
-
r
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
-
as active as trans-decenoyl-CoA
-
-
?
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
-
i.e. (E)-2-butenoyl-CoA. Reaction is catalyzed with a stereospecificity of 1 in 400000. The enzyme catalyzes the rapid interconversion of substrate and the (3S)-3-hydroxybutanoyl-CoA product relative to the rate of (3R)-3-hydroxybutanoyl-CoA formation. Formation of the correct product enantiomer requires an intact oxyanion hole and optimal positioning of the substrate with respect to two catalytic glutamates (E144 and E164) in the active site
-
-
r
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
-
i.e. (E)-2-butenoyl-CoA. The reaction proceeds via the syn addition of water and thus the pro-2R proton of (3S)-hydroxybutyryl-CoA is derived from solvent. The equilibrium constant for the hydration of trans-2-crotonyl-CoA to (3S)-hydroxybutyryl-CoA is 7.5. The rate of 3(R)-hydroxybutyryl-CoA formation is 400000fold slower than the normal hydration reaction (of crotonyl-CoA to (3S)-3-hydroxybutanoyl-CoA) but at least 1600000fold faster than the non-enzyme-catalyzed reaction. Formation of the incorrect stereoisomer likely occurs via syn addition of water to the incorrect face of the trans-2-crotonyl-CoA double bond. The absolute stereospecificity for the enzyme-catalyzed reaction is 1 in 400000
-
-
r
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
-
ratio of hydration rates trans-2-decenoyl-CoA/crotonyl-CoA is 0.29
-
-
r
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
stereoselective reaction mechanism, Glu144 and Glu164 are essential for ECH catalysis, overview
-
-
?
trans-2-decenoyl-CoA + H2O
(3S)-hydroxydecanoyl-CoA
-
ratio of hydration rates trans-2-decenoyl-CoA/crotonyl-CoA is 0.29
-
-
r
additional information
?
-
ECH catalyzes the reversible syn-addition of a water molecule across the double bond of a trans-2-enoyl-CoA, e.g. crotonyl-CoA, thioester to give a beta-hydroxyacyl-CoA thioester. The enzyme binds the substrates at the interface between monomers within the same trimer
-
-
?
additional information
?
-
-
In eukaryotes, ECH2 is a 31 kDa integral part of multifunctional protein-2, MFP-2, also called multifunctional enzyme 2, D-bifunctional enzyme, or 17-beta-estradiol dehydrogenase type IV. The MFP-2 plays a central role in peroxisomal beta-oxidation as it handles most peroxisomal beta-oxidation substrates
-
-
?
additional information
?
-
-
the beta-oxidation in mitochondria involves a (3S)-hydroxyacyl-CoA intermediate, while the beta-oxidation in peroxisomes has a (3R)-hydroxyacyl-CoA intermediate. The enzymes responsible for the formation of these two different intermediates are enoyl-CoA hydratase 1 (ECH1) in mitochondria and enoyl-CoA hydratase 2 (ECH2) in peroxisomes
-
-
?
additional information
?
-
ECH (XI) also has enoyl-CoA isomerase activity at approximately 1/5000 the level of its hydratase activity, overview
-
-
?
additional information
?
-
-
the enzyme also catalyzes DELTA3-DELTA2-isomerization of trans-3-hexenoyl-CoA
-
-
?
additional information
?
-
-
(3R)-3-hydroxyacyl-CoA is a peroxisomal specific intermediate
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-
?
additional information
?
-
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development and evaluation of a quantitative product separation method by a chiral column chromatography, overview
-
-
?
additional information
?
-
enzyme ECH displays activity toward unsaturated CoA thioesters with different chain lengths, although the turnover rate decreases for longer substrates
-
-
?
additional information
?
-
enzyme ECH displays activity toward unsaturated CoA thioesters with different chain lengths, although the turnover rate decreases for longer substrates
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(Z)-2-butenoyl-CoA + H2O
(3R)-3-hydroxybutanoyl-CoA
-
kcat is 12fold slower than with the trans-iosmer crotonyl-CoA
-
-
?
crotonyl-CoA + H2O
(3S)-3-hydroxybutanoyl-CoA
-
i.e. (E)-2-butenoyl-CoA. The reaction proceeds via the syn addition of water and thus the pro-2R proton of (3S)-hydroxybutyryl-CoA is derived from solvent. The equilibrium constant for the hydration of trans-2-crotonyl-CoA to (3S)-hydroxybutyryl-CoA is 7.5. The rate of 3(R)-hydroxybutyryl-CoA formation is 400000fold slower than the normal hydration reaction (of crotonyl-CoA to (3S)-3-hydroxybutanoyl-CoA) but at least 1600000fold faster than the non-enzyme-catalyzed reaction. Formation of the incorrect stereoisomer likely occurs via syn addition of water to the incorrect face of the trans-2-crotonyl-CoA double bond. The absolute stereospecificity for the enzyme-catalyzed reaction is 1 in 400000. To account for the exchange of the hydroxybutyryl pro-2S proton, the enzyme must also catalyze the dehydration of 3(R)-hydroxybutyryl-CoA to cis-2-crotonyl-CoA. Thus, the enzyme is capable of catalyzing the epimerization of hydroxybutyryl-CoA
-
-
r
methacrylyl-CoA + H2O
3-hydroxy-2-methylpropanoyl-CoA
-
-
-
r
additional information
?
-
ECH catalyzes the reversible syn-addition of a water molecule across the double bond of a trans-2-enoyl-CoA, e.g. crotonyl-CoA, thioester to give a beta-hydroxyacyl-CoA thioester. The enzyme binds the substrates at the interface between monomers within the same trimer
-
-
?
additional information
?
-
-
In eukaryotes, ECH2 is a 31 kDa integral part of multifunctional protein-2, MFP-2, also called multifunctional enzyme 2, D-bifunctional enzyme, or 17-beta-estradiol dehydrogenase type IV. The MFP-2 plays a central role in peroxisomal beta-oxidation as it handles most peroxisomal beta-oxidation substrates
-
-
?
additional information
?
-
-
the beta-oxidation in mitochondria involves a (3S)-hydroxyacyl-CoA intermediate, while the beta-oxidation in peroxisomes has a (3R)-hydroxyacyl-CoA intermediate. The enzymes responsible for the formation of these two different intermediates are enoyl-CoA hydratase 1 (ECH1) in mitochondria and enoyl-CoA hydratase 2 (ECH2) in peroxisomes
-
-
?
additional information
?
-
-
(3R)-3-hydroxyacyl-CoA is a peroxisomal specific intermediate
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(R)-methylenecyclopropylformyl-CoA
-
methylenecyclopropylformyl-CoA is a better inhibitor for enoyl-CoA hydratase 2 than for enoyl-CoA hydratase 1
(S)-methylenecyclopropylformyl-CoA
-
methylenecyclopropylformyl-CoA is a better inhibitor for enoyl-CoA hydratase 2 than for enoyl-CoA hydratase 1
3-octynoyl-CoA
-
irreversibly inactivates only enoyl-CoA hydratase 2, the catalytic residue Glu47 is covalently labeled by the inhibitor
methylenecyclopropylformyl-CoA
-
a metabolite derived from a natural amino acid, (methylenecyclopropyl)glycine, that inactivates enoyl-CoA hydratase 1 and enoyl-CoA hydratase 2. Competence of (R)- and (S)-MCPF-CoA to inactivate the ECH2 and kinetic analysis, enzyme-inhibitor complex structures, mass spectrometric analysis, inhibition mechanism, overview
additional information
-
3-octynoyl-CoA is not an inhibitor of enoyl-CoA hydratase 1
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
the classification is ambiguous because the stereochemistry is not exactly determined
UniProt
the classification is ambiguous because the stereochemistry is not exactly determined
-
-
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
malfunction
while mutation of Glu144 to alanine in this enzyme diminishes the isomerase activity by 10fold, mutation of Glu164 to alanine decreases the isomerase activity 1000fold, the hydratase activity is decreased 2000fold for both mutants
metabolism
the prototypical crotonases enoyl-CoA hydratase (ECH) and enoyl-CoA isomerase (ECI) are crucially involved in the beta-oxidation pathway of fatty acid metabolism
physiological function
evolution
the crotonases comprise a widely distributed enzyme superfamily that has multiple roles in both primary and secondary metabolism. Enoyl-CoA hydratase (ECH) and enoyl-CoA isomerase (ECI) are prototypical crotonases. The term crotonase has been used to refer specifically to ECH, but it is also used to refer to the entirety of the superfamily of enzymes bearing the crotonase-type fold. Some enzymes (e.g. rat peroxisomal multifunctional enzyme, type 1) have both ECH and ECI activities. These enzymes employ an active site with two glutamate residues. Rat mitochondrial ECH-1 (which has the two glutamate residues typical of ECH) has isomerase activity, albeit much lower than its hydratase activity. While the hydratase activity depends on both glutamate residues, the isomerase activity (as with dedicated ECI enzymes) relies mostly on a single glutamate
evolution
the crotonases comprise a widely distributed enzyme superfamily that has multiple roles in both primary and secondary metabolism. Enoyl-CoA hydratase (ECH) and enoyl-CoA isomerase (ECI) are prototypical crotonases. The term crotonase has been used to refer specifically to ECH, but it is also used to refer to the entirety of the superfamily of enzymes bearing the crotonase-type fold. Some enzymes, e.g. rat peroxisomal multifunctional enzyme, type 1, have both ECH and ECI activities. These enzymes employ an active site with two glutamate residues. Through the use of an additional domain, some multifunctional crotonase enzymes can also catalyze a further step in fatty acid catabolism, i.e. the oxidation of the enoyl-CoA hydratase product. While the hydratase activity depends on both glutamate residues, the isomerase activity (as with dedicated ECI enzymes) relies mostly on a single glutamate
physiological function
the multifunctional enzyme is involved in an alpha-methylacyl-CoAracemase-MFE2 independent synthesis pathway of bile acids from (24S)-hydroxyoxisterols, is involved in the beta-oxidation of long chain dicarboxylic acids
physiological function
prototypical crotonase enoyl-CoA hydratase (ECH) and enoyl-CoA isomerase (ECI) are crucially involved in the beta-oxidation pathway of fatty acid metabolism. Enzyme ECH catalyzes the second step of the beta-oxidation pathway: i.e. the syn addition of a water molecule across the double bond of an alpha,beta-unsaturated enoyl-CoA thioester substrate, e.g. crotonyl or methacrylyl-CoA
physiological function
prototypical crotonase enoyl-CoA hydratase (ECH) is crucially involved in the beta-oxidation pathway of fatty acid metabolism. Enzyme ECH catalyzes the second step of the beta-oxidation pathway: i.e. the syn addition of a water molecule across the double bond of an alpha,beta-unsaturated enoyl-CoA thioester substrate, e.g. crotonyl or methacrylyl-CoA. Rat mitochondrial ECH-1 (which has the two glutamate residues typical of ECH) has isomerase activity, albeit much lower than its hydratase activity
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). ECHA_RAT
763
0
82665
Swiss-Prot
Mitochondrion (Reliability: 1)
ECHM_RAT
290
0
31516
Swiss-Prot
Mitochondrion (Reliability: 1)
ECHP_RAT
722
0
78658
Swiss-Prot
other Location (Reliability: 4)
A0A8I6A8P4_RAT
718
0
78143
TrEMBL
Mitochondrion (Reliability: 1)
A0A8I5ZNC3_RAT
673
0
73537
TrEMBL
Mitochondrion (Reliability: 5)
ECH1_RAT
327
0
36172
Swiss-Prot
other Location (Reliability: 2)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexamer
homohexamer
the ECH structure adopts a functional hexamer comprising two stacked trimers
additional information
additional information
perMFE-1 can be divided into ®ve separate domains or parts
additional information
the enzyme binds the substrates at the interface between monomers within the same trimer, monomer structure, modelling in comparison to other crotonase superfamily enzymes, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
at 2.8 A resolution, multidomain protein having 5 domains: A, B, C, D, and E. The N-terminal part has a crotonase fold, which builds the active site for the DELTA3,DELTA2-enoyl-CoA isomerase (EC 5.3.3.8) and DELTA2-enoyl-CoA hydratase-1
hanging drop method, crystal structure of the enzyme complexed with the potent inhibitor acetoacetyl-CoA, refined at 2.5 A resolution. The active site architecture confirms the importance of Glu164 as the catalytic acid for providing the alpha-proton during the hydratase reaction. It also shows the importance of Glu144 as the catalytic base for the activation of a water molecule in the hydratase reaction
hanging drop method, structure of the mitochondrial enoyl-CoA hydratase, co-crystallised with the inhibitor octanoyl-CoA, refined at a resolution of 2.4 A
structure of enoyl-Coenzyme A (CoA) hydratase, co-crystallised with the inhibitor octanoyl-CoA, refined at a resolution of 2.4 A
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A98P
-
kcat is decreased 3400fold compared to wild type and KM is increased 13fold, mutant enzyme has a severely compromised ability for catalyzing the formation of (3R)-3-hydroxybutanoyl-CoA
E144A
E144A/Q162L
-
kcat for trans-2-hexenoyl-CoA is 12417fold lower than wild-type value. The point mutations E144A and Q162L by themselves apparently do not cause structural rearrangements of the active site helix, but when both residues are changed, the active site geometry changes
E144Q
-
3000fold decreases in kcat with little change in KM. The mutant is unable to catalyze the formation of (3R)-3-hydroxybutanoyl-CoA even when the incubation is extended to 4 days
E164A
E164D
-
1200fold decreases in kcat with little change in KM. First-order rate constant for the formation of (3R)-3-hydroxybutanoyl-CoA is similar to wild-type value
E164Q
-
340000fold decreases in kcat with little change in KM. While wild-type enoyl-CoA hydratase catalyzes the rapid interconversion of substrate and the (3S)-3-hydroxybutanoyl-CoA product relative to the rate of (3R)-3-hydroxybutanoyl-CoA formation, E164Q catalyzes the formation of both product enantiomers at similar rates
G141P
-
1600000fold decrease in kcat with no change in KM, mutant enzyme has a severely compromised ability for catalyzing the formation of (3R)-3-hydroxybutanoyl-CoA
Q162A
-
kcat for trans-2-hexenoyl-CoA is nearly identical to wild-type value
Q162L
-
kcat for trans-2-hexenoyl-CoA is nearly identical to wild-type value
Q162M
-
kcat for trans-2-hexenoyl-CoA is nearly identical to wild-type value
additional information
experiments with engineered perMFE-1 variants demonstrate that the H1/I competence of domain A requires stabilizing interactions with domains D and E. The variant His-perMFE (residues 288-79)DELTA, in which the domain C is deleted, is stable and has hydratase-1 activity
E144A
-
kcat for trans-2-hexenoyl-CoA is 1733fold lower than wild-type value
E144A
site-directed mutagenesis, the mutant shows a 1000fold reduced activity compared to the wild-type enzyme
E164A
-
kcat for trans-2-hexenoyl-CoA is 1709fold lower than wild-type value
E164A
site-directed mutagenesis, the mutant shows a 1000fold reduced activity compared to the wild-type enzyme
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
60
-
liver cell homogenate, 2 min, production of (3R)-3-hydroxyhexadecanoyl-CoA is remarkably decreased after heat treatment, while production of (3S)-3-hydroxyhexadecanoyl-CoA remains stable
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Pichia pastoris
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
a proteomic approach is applied to examine the effect of high fat diet on the liver proteome during the progression of nonalcoholic fatty liver disease. Male rats fed an high-fat diet for 4, 12, and 24 weeks show a reduced protein level of ECHS1
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
-
ECHS1 down-regulation contributes to high-fat diet-induced hepatic steatosis
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Engel, C.K.; Kiema, T.R.; Hiltunen, J.K.; Wierenga, R.K.
The crystal structure of enoyl-CoA hydratase complexed with octanoyl-CoA reveals the structural adaptations required for binding of a long chain fatty acid-CoA molecule
J. Mol. Biol.
275
847-859
1998
Rattus norvegicus (P14604)
Kiema, T.R.; Taskinen, J.P.; Pirilae, P.L.; Koivuranta, K.T.; Wierenga, R.K.; Hiltunen, J.K.
Organization of the multifunctional enzyme type 1: interaction between N- and C-terminal domains is required for the hydratase-1/isomerase activity
Biochem. J.
367
433-441
2002
Rattus norvegicus (P14604)
Kiema, T.R.; Engel, C.K.; Schmitz, W.; Filppula, S.A.; Wierenga, R.K.; Hiltunen, J.K.
Mutagenic and enzymological studies of the hydratase and isomerase activities of 2-enoyl-CoA hydratase-1
Biochemistry
38
2991-2999
1999
Rattus norvegicus
Feng, Y.; Hofstein, H.A.; Zwahlen, J.; Tonge, P.J.
Effect of mutagenesis on the stereochemistry of enoyl-CoA hydratase
Biochemistry
41
12883-12890
2002
Rattus norvegicus
Wu, W.J.; Feng, Y.; He, X.; Hofstein, H.A.; Raleigh, D.P.; Tonge, P.J.
Stereospecificity of the reaction catalyzed by enoyl-CoA hydratase
J. Am. Chem. Soc.
122
3987-3994
2000
Rattus norvegicus
-
Qin, Y.; Haapalainen, A.M.; Conry, D.; Cuebas, D.A.; Hiltunen, J.K.; Novikov, D.K.
Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis
Biochem. J.
328
377-382
1997
Rattus norvegicus
-
Hiltunen, J.K.; Palosaari, P.M.; Kunau, W.H.
Epimerization of 3-hydroxyacyl-CoA esters in rat liver. Involvement of two 2-enoyl-CoA hydratases
J. Biol. Chem.
264
13536-13540
1989
Rattus norvegicus
Wu, L.; Lin, S.; Li, D.
Comparative inhibition studies of enoyl-CoA hydratase 1 and enoyl-CoA hydratase 2 in long-chain fatty acid oxidation
Org. Lett.
10
3355-3358
2008
Rattus norvegicus
Qin, Y.M.; Poutanen, M.H.; Helander, H.M.; Kvist, A.P.; Siivari, K.M.; Schmitz, W.; Conzelmann, E.; Hellman, U.; Hiltunen, J.K.
Peroxisomal multifunctional enzyme of beta-oxidation metabolizing D-3-hydroxyacyl-CoA esters in rat liver: molecular cloning, expression and characterization
Biochem. J.
321
21-28
1997
Rattus norvegicus
Hamed, R.B.; Batchelar, E.T.; Clifton, I.J.; Schofield, C.J.
Mechanisms and structures of crotonase superfamily enzymes - how nature controls enolate and oxyanion reactivity
Cell. Mol. Life Sci.
65
2507-2527
2008
Rattus norvegicus (P14604)
Engel, C.K.; Mathieu, M.; Zeelen, J.P.; Hiltunen, J.K.; Wierenga, R.K.
Crystal structure of enoyl-coenzyme A (CoA) hydratase at 2.5 angstroms resolution: a spiral fold defines the CoA-binding pocket
EMBO J.
15
5135-5145
1996
Rattus norvegicus (P14604)
Minami-Ishii, N.; Taketani, S.; Osumi, T.; Hashimoto, T.
Molecular cloning and sequence analysis of the cDNA for rat mitochondrial enoyl-CoA hydratase. Structural and evolutionary relationships linked to the bifunctional enzyme of the peroxisomal beta-oxidation system
Eur. J. Biochem.
185
73-78
1989
Rattus norvegicus (P14604)
Engel, C.K.; Kiema, T.R.; Hiltunen, J.K.; Wierenga, R.K.
The crystal structure of enoyl-CoA hydratase complexed with octanoyl-CoA reveals the structural adaptations required for binding of a long chain fatty acid-CoA molecule
J. Mol. Biol.
275
847-859
1998
Rattus norvegicus
Hiromasa, Y.; Yan, X.; Roche, T.E.
Specific ion influences on self-association of pyruvate dehydrogenase kinase isoform 2 (PDHK2), binding of PDHK2 to the L2 lipoyl domain, and effects of the lipoyl group-binding site inhibitor, Nov3r
Biochemistry
47
2312-2324
2008
Homo sapiens, Mus musculus, Rattus norvegicus
Kasaragod, P.; Venkatesan, R.; Kiema, T.R.; Hiltunen, J.K.; Wierenga, R.K.
The crystal structure of liganded rat peroxisomal multifunctional enzyme type 1: a flexible molecule with two interconnected active sites
J. Biol. Chem.
285
24089-24098
2010
Rattus norvegicus (P07896)
Tsuchida, S.; Kawamoto, K.; Nunome, K.; Hamaue, N.; Yoshimura, T.; Aoki, T.; Kurosawa, T.
Analysis of enoyl-coenzyme A hydratase activity and its stereospecificity using high-performance liquid chromatography equipped with chiral separation column
J. Oleo Sci.
60
221-228
2011
Rattus norvegicus
Lohans, C.; Wang, D.; Wang, J.; Hamed, R.; Schofield, C.
Crotonases natures exceedingly convertible catalysts
ACS Catal.
7
6587-6599
2017
Streptomyces sp. V-1, Rattus norvegicus (P07896), Rattus norvegicus (P14604)
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