ECHM_RAT
ID ECHM_RAT Reviewed; 290 AA.
AC P14604;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-APR-1990, sequence version 1.
DT 03-AUG-2022, entry version 178.
DE RecName: Full=Enoyl-CoA hydratase, mitochondrial {ECO:0000305};
DE Short=mECH {ECO:0000303|PubMed:7883013};
DE Short=mECH1 {ECO:0000303|PubMed:7883013};
DE EC=4.2.1.17 {ECO:0000269|PubMed:10074351, ECO:0000269|PubMed:7883013};
DE EC=5.3.3.8 {ECO:0000269|PubMed:10074351};
DE AltName: Full=Enoyl-CoA hydratase 1;
DE Short=ECHS1;
DE AltName: Full=Short-chain enoyl-CoA hydratase;
DE Short=SCEH;
DE Flags: Precursor;
GN Name=Echs1 {ECO:0000312|RGD:69330};
OS Rattus norvegicus (Rat).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC Murinae; Rattus.
OX NCBI_TaxID=10116;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], AND PROTEIN SEQUENCE OF 30-39 AND 80-94.
RC TISSUE=Liver;
RX PubMed=2806264; DOI=10.1111/j.1432-1033.1989.tb15083.x;
RA Minami-Ishii N., Taketani S., Osumi T., Hashimoto T.;
RT "Molecular cloning and sequence analysis of the cDNA for rat mitochondrial
RT enoyl-CoA hydratase. Structural and evolutionary relationships linked to
RT the bifunctional enzyme of the peroxisomal beta-oxidation system.";
RL Eur. J. Biochem. 185:73-78(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC TISSUE=Prostate;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA project:
RT the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [3]
RP MUTAGENESIS OF GLU-164, CATALYTIC ACTIVITY, AND FUNCTION.
RX PubMed=7883013; DOI=10.1111/j.1432-1033.1995.tb20230.x;
RA Mueller-Newen G., Janssen U., Stoffel W.;
RT "Enoyl-CoA hydratase and isomerase form a superfamily with a common active-
RT site glutamate residue.";
RL Eur. J. Biochem. 228:68-73(1995).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=10074351; DOI=10.1021/bi981646v;
RA Kiema T.R., Engel C.K., Schmitz W., Filppula S.A., Wierenga R.K.,
RA Hiltunen J.K.;
RT "Mutagenic and enzymological studies of the hydratase and isomerase
RT activities of 2-enoyl-CoA hydratase-1.";
RL Biochemistry 38:2991-2999(1999).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH OCTANOYL COENZYME A,
RP AND SUBUNIT.
RC TISSUE=Liver;
RX PubMed=8895557; DOI=10.1002/j.1460-2075.1996.tb00897.x;
RA Engel C.K., Mathieu M., Zeelen J.P., Hiltunen J.K., Wierenga R.K.;
RT "Crystal structure of enoyl-coenzyme A (CoA) hydratase at 2.5-A resolution:
RT a spiral fold defines the CoA-binding pocket.";
RL EMBO J. 15:5135-5145(1996).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) IN COMPLEX WITH ACETOACETYL COENZYME
RP A, SUBUNIT, SUBCELLULAR LOCATION, AND TISSUE SPECIFICITY.
RX PubMed=9480773; DOI=10.1006/jmbi.1997.1491;
RA Engel C.K., Kiema T.R., Hiltunen J.K., Wierenga R.K.;
RT "The crystal structure of enoyl-CoA hydratase complexed with octanoyl-CoA
RT reveals the structural adaptations required for binding of a long chain
RT fatty acid-CoA molecule.";
RL J. Mol. Biol. 275:847-859(1998).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF 33-290 IN COMPLEX WITH
RP 4-N,N-DIMETHYLAMINOCINNAMOYL COENZYME A, ENZYME MECHANISM, AND SUBUNIT.
RX PubMed=11851409; DOI=10.1021/bi015844p;
RA Bahnson B.J., Anderson V.E., Petsko G.A.;
RT "Structural mechanism of enoyl-CoA hydratase: three atoms from a single
RT water are added in either an E1cb stepwise or concerted fashion.";
RL Biochemistry 41:2621-2629(2002).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 31-290 IN COMPLEX WITH HEXADIENOL
RP COENZYME A, AND MUTAGENESIS OF GLU-144.
RX PubMed=12445775; DOI=10.1016/s1074-5521(02)00263-6;
RA Bell A.F., Feng Y., Hofstein H.A., Parikh S., Wu J., Rudolph M.J.,
RA Kisker C., Whitty A., Tonge P.J.;
RT "Stereoselectivity of enoyl-CoA hydratase results from preferential
RT activation of one of two bound substrate conformers.";
RL Chem. Biol. 9:1247-1255(2002).
CC -!- FUNCTION: Converts unsaturated trans-2-enoyl-CoA species ((2E)-enoyl-
CC CoA) to the corresponding 3(S)-3-hydroxyacyl-CoA species through
CC addition of a water molecule to the double bond (PubMed:7883013,
CC PubMed:10074351). Catalyzes the hydration of medium- and short-chained
CC fatty enoyl-CoA thioesters from 4 carbons long (C4) up to C16
CC (PubMed:7883013, PubMed:10074351). Has high substrate specificity for
CC crotonyl-CoA ((2E)-butenoyl-CoA) and moderate specificity for acryloyl-
CC CoA, 3-methylcrotonyl-CoA (3-methyl-(2E)-butenoyl-CoA) and methacrylyl-
CC CoA ((2E)-2-methylpropenoyl-CoA). Can bind tiglyl-CoA (2-
CC methylcrotonoyl-CoA), but hydrates only a small amount of this
CC substrate (By similarity). Plays a key role in the beta-oxidation
CC spiral of short- and medium-chain fatty acid oxidation
CC (PubMed:7883013). At a lower rate than the hydratase reaction,
CC catalyzes the isomerase reaction of trans-3-enoyl-CoA species (such as
CC (3E)-hexenoyl-CoA) to trans-2-enoyl-CoA species (such as (2E)-hexenoyl-
CC CoA), which are subsequently hydrated to 3(S)-3-hydroxyacyl-CoA species
CC (such as (3S)-hydroxyhexanoyl-CoA) (PubMed:10074351).
CC {ECO:0000250|UniProtKB:P30084, ECO:0000269|PubMed:10074351,
CC ECO:0000269|PubMed:7883013}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a (3S)-3-hydroxyacyl-CoA = a (2E)-enoyl-CoA + H2O;
CC Xref=Rhea:RHEA:16105, ChEBI:CHEBI:15377, ChEBI:CHEBI:57318,
CC ChEBI:CHEBI:58856; EC=4.2.1.17;
CC Evidence={ECO:0000269|PubMed:10074351, ECO:0000269|PubMed:7883013};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:16107;
CC Evidence={ECO:0000305|PubMed:10074351, ECO:0000305|PubMed:7883013};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a (3E)-enoyl-CoA = a 4-saturated (2E)-enoyl-CoA;
CC Xref=Rhea:RHEA:45228, ChEBI:CHEBI:58521, ChEBI:CHEBI:85097;
CC EC=5.3.3.8; Evidence={ECO:0000269|PubMed:10074351};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:45229;
CC Evidence={ECO:0000305|PubMed:10074351};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(3E)-hexenoyl-CoA = (2E)-hexenoyl-CoA; Xref=Rhea:RHEA:45736,
CC ChEBI:CHEBI:62077, ChEBI:CHEBI:84790;
CC Evidence={ECO:0000269|PubMed:10074351};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:45737;
CC Evidence={ECO:0000305|PubMed:10074351};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(3S)-3-hydroxybutanoyl-CoA = (2E)-butenoyl-CoA + H2O;
CC Xref=Rhea:RHEA:26558, ChEBI:CHEBI:15377, ChEBI:CHEBI:57316,
CC ChEBI:CHEBI:57332; Evidence={ECO:0000269|PubMed:7883013};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:26560;
CC Evidence={ECO:0000305|PubMed:7883013};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-hydroxyisovaleryl-CoA = 3-methyl-(2E)-butenoyl-CoA + H2O;
CC Xref=Rhea:RHEA:31079, ChEBI:CHEBI:15377, ChEBI:CHEBI:57344,
CC ChEBI:CHEBI:62555; Evidence={ECO:0000250|UniProtKB:P30084};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:31081;
CC Evidence={ECO:0000250|UniProtKB:P30084};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-hydroxypropanoyl-CoA = acryloyl-CoA + H2O;
CC Xref=Rhea:RHEA:26518, ChEBI:CHEBI:15377, ChEBI:CHEBI:57367,
CC ChEBI:CHEBI:58528; Evidence={ECO:0000250|UniProtKB:P30084};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:26520;
CC Evidence={ECO:0000250|UniProtKB:P30084};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-hydroxybutanoyl-CoA = (2E)-butenoyl-CoA + H2O;
CC Xref=Rhea:RHEA:45584, ChEBI:CHEBI:15377, ChEBI:CHEBI:57332,
CC ChEBI:CHEBI:78611; Evidence={ECO:0000250|UniProtKB:P30084};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:45586;
CC Evidence={ECO:0000250|UniProtKB:P30084};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E)-2-methylpropenoyl-CoA + H2O = (S)-3-hydroxyisobutanoyl-
CC CoA; Xref=Rhea:RHEA:31175, ChEBI:CHEBI:15377, ChEBI:CHEBI:62500,
CC ChEBI:CHEBI:62611; Evidence={ECO:0000250|UniProtKB:P30084};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:31176;
CC Evidence={ECO:0000250|UniProtKB:P30084};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(3S)-hydroxyhexanoyl-CoA = (2E)-hexenoyl-CoA + H2O;
CC Xref=Rhea:RHEA:30547, ChEBI:CHEBI:15377, ChEBI:CHEBI:62075,
CC ChEBI:CHEBI:62077; Evidence={ECO:0000269|PubMed:10074351};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:30549;
CC Evidence={ECO:0000305|PubMed:10074351};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(3S)-hydroxydecanoyl-CoA = (2E)-decenoyl-CoA + H2O;
CC Xref=Rhea:RHEA:31191, ChEBI:CHEBI:15377, ChEBI:CHEBI:61406,
CC ChEBI:CHEBI:62616; Evidence={ECO:0000269|PubMed:10074351};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:31193;
CC Evidence={ECO:0000305|PubMed:10074351};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=25.0 uM for (2E)-hexenoyl-CoA {ECO:0000269|PubMed:10074351};
CC KM=4.9 uM for (2E)-decenoyl-CoA {ECO:0000269|PubMed:10074351};
CC KM=65.5 uM for (3E)-hexenoyl-CoA {ECO:0000269|PubMed:10074351};
CC KM=49.9 uM for crotonyl-CoA ((2E)-butenoyl-CoA)
CC {ECO:0000269|PubMed:10074351};
CC Vmax=64 umol/min/mg enzyme using (2E)-hexenoyl-CoA as substrate
CC {ECO:0000269|PubMed:10074351};
CC -!- PATHWAY: Lipid metabolism; fatty acid beta-oxidation.
CC {ECO:0000250|UniProtKB:P30084}.
CC -!- SUBUNIT: Homohexamer; dimer of trimers. {ECO:0000269|PubMed:11851409,
CC ECO:0000269|PubMed:12445775, ECO:0000269|PubMed:8895557,
CC ECO:0000269|PubMed:9480773}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion matrix
CC {ECO:0000269|PubMed:9480773}.
CC -!- TISSUE SPECIFICITY: Detected in liver (at protein level).
CC {ECO:0000269|PubMed:9480773}.
CC -!- SIMILARITY: Belongs to the enoyl-CoA hydratase/isomerase family.
CC {ECO:0000305}.
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DR EMBL; X15958; CAA34080.1; -; mRNA.
DR EMBL; BC064655; AAH64655.1; -; mRNA.
DR PIR; S06477; S06477.
DR RefSeq; NP_511178.1; NM_078623.2.
DR RefSeq; XP_003748934.1; XM_003748886.3.
DR PDB; 1DUB; X-ray; 2.50 A; A/B/C/D/E/F=30-290.
DR PDB; 1EY3; X-ray; 2.30 A; A/B/C/D/E/F=33-290.
DR PDB; 1MJ3; X-ray; 2.10 A; A/B/C/D/E/F=31-290.
DR PDB; 2DUB; X-ray; 2.40 A; A/B/C/D/E/F=30-290.
DR PDBsum; 1DUB; -.
DR PDBsum; 1EY3; -.
DR PDBsum; 1MJ3; -.
DR PDBsum; 2DUB; -.
DR AlphaFoldDB; P14604; -.
DR SMR; P14604; -.
DR BioGRID; 250789; 1.
DR IntAct; P14604; 9.
DR STRING; 10116.ENSRNOP00000025446; -.
DR ChEMBL; CHEMBL3153; -.
DR SwissLipids; SLP:000001128; -.
DR iPTMnet; P14604; -.
DR PhosphoSitePlus; P14604; -.
DR jPOST; P14604; -.
DR PaxDb; P14604; -.
DR PRIDE; P14604; -.
DR Ensembl; ENSRNOT00000071574; ENSRNOP00000066388; ENSRNOG00000064647.
DR GeneID; 140547; -.
DR KEGG; rno:140547; -.
DR UCSC; RGD:69330; rat.
DR CTD; 1892; -.
DR RGD; 69330; Echs1.
DR eggNOG; KOG1680; Eukaryota.
DR GeneTree; ENSGT00940000157609; -.
DR HOGENOM; CLU_009834_7_6_1; -.
DR InParanoid; P14604; -.
DR OMA; IFKQTDA; -.
DR OrthoDB; 1221604at2759; -.
DR PhylomeDB; P14604; -.
DR TreeFam; TF314497; -.
DR BRENDA; 4.2.1.17; 5301.
DR BRENDA; 5.3.3.21; 5301.
DR Reactome; R-RNO-70895; Branched-chain amino acid catabolism.
DR Reactome; R-RNO-77310; Beta oxidation of lauroyl-CoA to decanoyl-CoA-CoA.
DR Reactome; R-RNO-77346; Beta oxidation of decanoyl-CoA to octanoyl-CoA-CoA.
DR Reactome; R-RNO-77348; Beta oxidation of octanoyl-CoA to hexanoyl-CoA.
DR Reactome; R-RNO-77350; Beta oxidation of hexanoyl-CoA to butanoyl-CoA.
DR Reactome; R-RNO-77352; Beta oxidation of butanoyl-CoA to acetyl-CoA.
DR UniPathway; UPA00659; -.
DR EvolutionaryTrace; P14604; -.
DR PRO; PR:P14604; -.
DR Proteomes; UP000002494; Chromosome 1.
DR Bgee; ENSRNOG00000018522; Expressed in heart and 19 other tissues.
DR Genevisible; P14604; RN.
DR GO; GO:0005759; C:mitochondrial matrix; IEA:UniProtKB-SubCell.
DR GO; GO:0005739; C:mitochondrion; IDA:RGD.
DR GO; GO:0043956; F:3-hydroxypropionyl-CoA dehydratase activity; IEA:RHEA.
DR GO; GO:0120092; F:crotonyl-CoA hydratase activity; IEA:RHEA.
DR GO; GO:0004165; F:delta(3)-delta(2)-enoyl-CoA isomerase activity; IEA:RHEA.
DR GO; GO:0004300; F:enoyl-CoA hydratase activity; IDA:RGD.
DR GO; GO:0006635; P:fatty acid beta-oxidation; IMP:RGD.
DR Gene3D; 1.10.12.10; -; 1.
DR InterPro; IPR029045; ClpP/crotonase-like_dom_sf.
DR InterPro; IPR018376; Enoyl-CoA_hyd/isom_CS.
DR InterPro; IPR001753; Enoyl-CoA_hydra/iso.
DR InterPro; IPR014748; Enoyl-CoA_hydra_C.
DR Pfam; PF00378; ECH_1; 1.
DR SUPFAM; SSF52096; SSF52096; 1.
DR PROSITE; PS00166; ENOYL_COA_HYDRATASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylation; Direct protein sequencing;
KW Fatty acid metabolism; Isomerase; Lipid metabolism; Lyase; Mitochondrion;
KW Reference proteome; Transit peptide.
FT TRANSIT 1..29
FT /note="Mitochondrion"
FT /evidence="ECO:0000269|PubMed:2806264"
FT CHAIN 30..290
FT /note="Enoyl-CoA hydratase, mitochondrial"
FT /id="PRO_0000007414"
FT BINDING 98..101
FT /ligand="substrate"
FT BINDING 141
FT /ligand="substrate"
FT SITE 164
FT /note="Important for catalytic activity"
FT MOD_RES 101
FT /note="N6-acetyllysine; alternate"
FT /evidence="ECO:0000250|UniProtKB:Q8BH95"
FT MOD_RES 101
FT /note="N6-succinyllysine; alternate"
FT /evidence="ECO:0000250|UniProtKB:Q8BH95"
FT MOD_RES 115
FT /note="N6-acetyllysine; alternate"
FT /evidence="ECO:0000250|UniProtKB:Q8BH95"
FT MOD_RES 115
FT /note="N6-succinyllysine; alternate"
FT /evidence="ECO:0000250|UniProtKB:Q8BH95"
FT MOD_RES 204
FT /note="N6-succinyllysine"
FT /evidence="ECO:0000250|UniProtKB:Q8BH95"
FT MOD_RES 211
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:Q8BH95"
FT MUTAGEN 144
FT /note="E->D: Reduces activity 50-fold."
FT /evidence="ECO:0000269|PubMed:12445775"
FT MUTAGEN 144
FT /note="E->Q: Reduces activity 3300-fold."
FT /evidence="ECO:0000269|PubMed:12445775"
FT MUTAGEN 164
FT /note="E->D: Reduces activity 1250-fold."
FT /evidence="ECO:0000269|PubMed:7883013"
FT MUTAGEN 164
FT /note="E->Q: Reduces activity 330000-fold."
FT /evidence="ECO:0000269|PubMed:7883013"
FT STRAND 34..42
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 43..45
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 47..52
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 55..57
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 63..78
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 84..88
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 91..95
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 100..103
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 108..113
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 119..125
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 130..134
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 136..139
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 141..148
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 149..155
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 159..161
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 163..167
FT /evidence="ECO:0007829|PDB:1MJ3"
FT TURN 175..177
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 178..183
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 185..194
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 197..199
FT /evidence="ECO:0007829|PDB:2DUB"
FT HELIX 200..205
FT /evidence="ECO:0007829|PDB:1MJ3"
FT STRAND 210..213
FT /evidence="ECO:0007829|PDB:1MJ3"
FT TURN 215..217
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 218..231
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 234..245
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 246..248
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 252..265
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 266..268
FT /evidence="ECO:0007829|PDB:1MJ3"
FT HELIX 270..280
FT /evidence="ECO:0007829|PDB:1MJ3"
SQ SEQUENCE 290 AA; 31516 MW; 1E528590961D0CC0 CRC64;
MAALRALLPR ACNSLLSPVR CPEFRRFASG ANFQYIITEK KGKNSSVGLI QLNRPKALNA
LCNGLIEELN QALETFEEDP AVGAIVLTGG EKAFAAGADI KEMQNRTFQD CYSGKFLSHW
DHITRIKKPV IAAVNGYALG GGCELAMMCD IIYAGEKAQF GQPEILLGTI PGAGGTQRLT
RAVGKSLAME MVLTGDRISA QDAKQAGLVS KIFPVETLVE EAIQCAEKIA NNSKIIVAMA
KESVNAAFEM TLTEGNKLEK KLFYSTFATD DRREGMSAFV EKRKANFKDH
