ID   MHPC_ECOLI              Reviewed;         288 AA.
AC   P77044; P71204; P77205; Q2MC75;
DT   01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT   14-DEC-2011, sequence version 4.
DT   27-MAR-2024, entry version 169.
DE   RecName: Full=2-hydroxy-6-oxononadienedioate/2-hydroxy-6-oxononatrienedioate hydrolase {ECO:0000255|HAMAP-Rule:MF_01654};
DE            EC=3.7.1.14 {ECO:0000255|HAMAP-Rule:MF_01654, ECO:0000269|PubMed:15663941};
DE   AltName: Full=2-hydroxy-6-ketonona-2,4-diene-1,9-dioic acid 5,6-hydrolase {ECO:0000255|HAMAP-Rule:MF_01654};
DE   AltName: Full=2-hydroxy-6-oxonona-2,4,7-triene-1,9-dioic acid 5,6-hydrolase {ECO:0000255|HAMAP-Rule:MF_01654};
DE   AltName: Full=2-hydroxy-6-oxonona-2,4-diene-1,9-dioic acid 5,6-hydrolase {ECO:0000255|HAMAP-Rule:MF_01654};
GN   Name=mhpC {ECO:0000255|HAMAP-Rule:MF_01654};
GN   OrderedLocusNames=b0349, JW0340;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RA   Kawamukai M.;
RT   "Complete sequence of the mhp operon.";
RL   Submitted (JUN-1996) to the EMBL/GenBank/DDBJ databases.
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION AS A HYDROLASE.
RC   STRAIN=K12 / CS520;
RX   PubMed=9098055; DOI=10.1128/jb.179.8.2573-2581.1997;
RA   Ferrandez A., Garcia J.L., Diaz E.;
RT   "Genetic characterization and expression in heterologous hosts of the 3-(3-
RT   hydroxyphenyl)propionate catabolic pathway of Escherichia coli K-12.";
RL   J. Bacteriol. 179:2573-2581(1997).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RA   Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA   Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA   Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT   "Sequence of minutes 4-25 of Escherichia coli.";
RL   Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN   [4]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA   Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [5]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT   and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [6]
RP   PROTEIN SEQUENCE OF 2-18, FUNCTION AS A HYDROLASE, BIOPHYSICOCHEMICAL
RP   PROPERTIES, AND SUBUNIT.
RX   PubMed=9315862; DOI=10.1021/bi971115r;
RA   Lam W.W., Bugg T.D.;
RT   "Purification, characterization, and stereochemical analysis of a C-C
RT   hydrolase: 2-hydroxy-6-keto-nona-2,4-diene-1,9-dioic acid 5,6-hydrolase.";
RL   Biochemistry 36:12242-12251(1997).
RN   [7]
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP   SER-44; SER-114; HIS-118 AND HIS-267, ACTIVE SITE, AND REACTION MECHANISM.
RX   PubMed=15663941; DOI=10.1016/j.jmb.2004.11.032;
RA   Li C., Montgomery M.G., Mohammed F., Li J.-J., Wood S.P., Bugg T.D.H.;
RT   "Catalytic mechanism of C-C hydrolase MhpC from Escherichia coli: kinetic
RT   analysis of His263 and Ser110 site-directed mutants.";
RL   J. Mol. Biol. 346:241-251(2005).
RN   [8]
RP   MUTAGENESIS OF ASN-113; PHE-177; ARG-192; CYS-265 AND TRP-268, AND
RP   CATALYTIC MECHANISM.
RX   PubMed=17029402; DOI=10.1021/bi061253t;
RA   Li C., Li J.-J., Montgomery M.G., Wood S.P., Bugg T.D.H.;
RT   "Catalytic role for arginine 188 in the C-C hydrolase catalytic mechanism
RT   for Escherichia coli MhpC and Burkholderia xenovorans LB400 BphD.";
RL   Biochemistry 45:12470-12479(2006).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 5-288 IN COMPLEX WITH A
RP   NON-CLEAVABLE SUBSTRATE ANALOG, MASS SPECTROMETRY, AND SUBUNIT.
RX   PubMed=15663942; DOI=10.1016/j.jmb.2004.11.033;
RA   Dunn G., Montgomery M.G., Mohammed F., Coker A., Cooper J.B., Robertson T.,
RA   Garcia J.-L., Bugg T.D.H., Wood S.P.;
RT   "The structure of the C-C bond hydrolase MhpC provides insights into its
RT   catalytic mechanism.";
RL   J. Mol. Biol. 346:253-265(2005).
CC   -!- FUNCTION: Catalyzes the cleavage of the C5-C6 bond of 2-hydroxy-6-
CC       oxononadienedioate and 2-hydroxy-6-oxononatrienedioate, a dienol ring
CC       fission product of the bacterial meta-cleavage pathway for degradation
CC       of phenylpropionic acid. MhpC shows some selectivity for the
CC       carboxylate of the side chain. {ECO:0000269|PubMed:15663941,
CC       ECO:0000269|PubMed:9098055, ECO:0000269|PubMed:9315862}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(2Z,4E)-2-hydroxy-6-oxonona-2,4-dienedioate + H2O = (2Z)-2-
CC         hydroxypenta-2,4-dienoate + H(+) + succinate; Xref=Rhea:RHEA:34187,
CC         ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:30031,
CC         ChEBI:CHEBI:66887, ChEBI:CHEBI:67152; EC=3.7.1.14;
CC         Evidence={ECO:0000255|HAMAP-Rule:MF_01654,
CC         ECO:0000269|PubMed:15663941};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(2Z,4E,7E)-2-hydroxy-6-oxonona-2,4,7-trienedioate + H2O =
CC         (2Z)-2-hydroxypenta-2,4-dienoate + fumarate + H(+);
CC         Xref=Rhea:RHEA:34191, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:29806, ChEBI:CHEBI:66888, ChEBI:CHEBI:67152; EC=3.7.1.14;
CC         Evidence={ECO:0000255|HAMAP-Rule:MF_01654,
CC         ECO:0000269|PubMed:15663941};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=2.1 uM for 2-hydroxy-6-oxononadienedioate (at pH 8)
CC         {ECO:0000269|PubMed:9315862};
CC         KM=6.8 uM for 2-hydroxy-6-oxononadienedioate (at pH 8)
CC         {ECO:0000269|PubMed:15663941};
CC         Note=kcat is 28 sec(-1) for hydrolase activity with
CC         2-hydroxy-6-oxononadienedioate as substrate.
CC         {ECO:0000269|PubMed:15663941};
CC   -!- PATHWAY: Aromatic compound metabolism; 3-phenylpropanoate degradation.
CC       {ECO:0000255|HAMAP-Rule:MF_01654}.
CC   -!- SUBUNIT: Homodimer. {ECO:0000255|HAMAP-Rule:MF_01654,
CC       ECO:0000269|PubMed:15663942, ECO:0000269|PubMed:9315862}.
CC   -!- MASS SPECTROMETRY: Mass=31717; Method=Electrospray;
CC       Evidence={ECO:0000269|PubMed:15663942};
CC   -!- MISCELLANEOUS: MhpC is not a serine hydrolase (catalytic triad), as
CC       Ser-114 is a non-nucleophilic catalytic residue and Asp-239 is not
CC       involved in the catalytic mechanism.
CC   -!- SIMILARITY: Belongs to the AB hydrolase superfamily. MhpC family.
CC       {ECO:0000255|HAMAP-Rule:MF_01654, ECO:0000305}.
CC   -!- SEQUENCE CAUTION:
CC       Sequence=AAB18073.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
CC       Sequence=BAA13054.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
CC       Sequence=BAE76131.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
CC       Sequence=CAA70749.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR   EMBL; D86239; BAA13054.1; ALT_INIT; Genomic_DNA.
DR   EMBL; Y09555; CAA70749.1; ALT_INIT; Genomic_DNA.
DR   EMBL; U73857; AAB18073.1; ALT_INIT; Genomic_DNA.
DR   EMBL; U00096; AAC73452.3; -; Genomic_DNA.
DR   EMBL; AP009048; BAE76131.1; ALT_INIT; Genomic_DNA.
DR   PIR; E64762; E64762.
DR   RefSeq; NP_414883.5; NC_000913.3.
DR   RefSeq; WP_000121907.1; NZ_SSZK01000061.1.
DR   PDB; 1U2E; X-ray; 2.10 A; A/B/C/D=1-288.
DR   PDBsum; 1U2E; -.
DR   AlphaFoldDB; P77044; -.
DR   SMR; P77044; -.
DR   BioGRID; 4260729; 8.
DR   DIP; DIP-10207N; -.
DR   IntAct; P77044; 3.
DR   STRING; 511145.b0349; -.
DR   SwissLipids; SLP:000001887; -.
DR   ESTHER; ecoli-mhpc; Carbon-carbon_bond_hydrolase.
DR   MEROPS; S33.995; -.
DR   PaxDb; 511145-b0349; -.
DR   EnsemblBacteria; AAC73452; AAC73452; b0349.
DR   GeneID; 944954; -.
DR   KEGG; ecj:JW0340; -.
DR   KEGG; eco:b0349; -.
DR   PATRIC; fig|1411691.4.peg.1929; -.
DR   EchoBASE; EB4168; -.
DR   eggNOG; COG0596; Bacteria.
DR   HOGENOM; CLU_020336_13_2_6; -.
DR   InParanoid; P77044; -.
DR   OrthoDB; 5853561at2; -.
DR   PhylomeDB; P77044; -.
DR   BioCyc; EcoCyc:MHPCHYDROL-MONOMER; -.
DR   BioCyc; MetaCyc:MHPCHYDROL-MONOMER; -.
DR   BRENDA; 3.7.1.14; 2026.
DR   UniPathway; UPA00714; -.
DR   EvolutionaryTrace; P77044; -.
DR   PRO; PR:P77044; -.
DR   Proteomes; UP000000318; Chromosome.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005737; C:cytoplasm; IDA:EcoliWiki.
DR   GO; GO:0052823; F:2-hydroxy-6-oxonona-2,4,7-trienedioate hydrolase activity; IDA:EcoCyc.
DR   GO; GO:0018771; F:2-hydroxy-6-oxonona-2,4-dienedioate hydrolase activity; IDA:EcoCyc.
DR   GO; GO:0016787; F:hydrolase activity; IDA:EcoliWiki.
DR   GO; GO:0042803; F:protein homodimerization activity; IEA:InterPro.
DR   GO; GO:0019622; P:3-(3-hydroxy)phenylpropionate catabolic process; IMP:EcoCyc.
DR   GO; GO:0019380; P:3-phenylpropionate catabolic process; IEA:UniProtKB-UniPathway.
DR   GO; GO:0019439; P:aromatic compound catabolic process; IMP:EcoliWiki.
DR   Gene3D; 3.40.50.1820; alpha/beta hydrolase; 1.
DR   HAMAP; MF_01654; MhpC; 1.
DR   InterPro; IPR029058; AB_hydrolase.
DR   InterPro; IPR000073; AB_hydrolase_1.
DR   InterPro; IPR000639; Epox_hydrolase-like.
DR   InterPro; IPR023791; MhpC_alpha/beta_hydrolase.
DR   PANTHER; PTHR43689:SF53; ALPHA_BETA-HYDROLASES SUPERFAMILY PROTEIN; 1.
DR   PANTHER; PTHR43689; HYDROLASE; 1.
DR   Pfam; PF00561; Abhydrolase_1; 1.
DR   PRINTS; PR00111; ABHYDROLASE.
DR   PRINTS; PR00412; EPOXHYDRLASE.
DR   SUPFAM; SSF53474; alpha/beta-Hydrolases; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Aromatic hydrocarbons catabolism; Direct protein sequencing;
KW   Hydrolase; Reference proteome.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:9315862"
FT   CHAIN           2..288
FT                   /note="2-hydroxy-6-oxononadienedioate/2-hydroxy-6-
FT                   oxononatrienedioate hydrolase"
FT                   /id="PRO_0000207054"
FT   ACT_SITE        267
FT                   /note="Proton acceptor"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_01654,
FT                   ECO:0000269|PubMed:15663941"
FT   SITE            114
FT                   /note="Transition state stabilizer"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_01654,
FT                   ECO:0000269|PubMed:15663941"
FT   SITE            192
FT                   /note="Catalytic role in ketonization of the dienol
FT                   substrate (substrate destabilization)"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_01654,
FT                   ECO:0000269|PubMed:15663941, ECO:0000269|PubMed:17029402"
FT   MUTAGEN         44
FT                   /note="S->A: 2-fold decrease in catalytic efficiency and
FT                   more than 5-fold increase in affinity for the natural
FT                   substrate."
FT                   /evidence="ECO:0000269|PubMed:15663941"
FT   MUTAGEN         113
FT                   /note="N->A: 200-fold decrease in catalytic activity and
FT                   almost 2-fold increase in affinity."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         113
FT                   /note="N->H: 350-fold decrease in catalytic activity and
FT                   almost 2-fold increase in affinity."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         114
FT                   /note="S->A: Weakly active. 3-fold decrease in affinity.
FT                   Fast ketonisation and slow C-C cleavage."
FT                   /evidence="ECO:0000269|PubMed:15663941"
FT   MUTAGEN         114
FT                   /note="S->G: Weakly active. 3-fold decrease in affinity.
FT                   Fast ketonisation and slow C-C cleavage."
FT                   /evidence="ECO:0000269|PubMed:15663941"
FT   MUTAGEN         118
FT                   /note="H->A: More than 2-fold decrease in catalytic
FT                   efficiency and 3-fold increase affinity."
FT                   /evidence="ECO:0000269|PubMed:15663941"
FT   MUTAGEN         177
FT                   /note="F->D: 100-fold decrease in catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         177
FT                   /note="F->G: 4-fold and 8-fold decrease in catalytic
FT                   activity and affinity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         192
FT                   /note="R->K: 40-fold and 5-fold decrease in catalytic
FT                   activity and affinity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         192
FT                   /note="R->Q: 280-fold and 10-fold decrease in catalytic
FT                   activity and affinity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         265
FT                   /note="C->A: 2-fold decrease in catalytic activity and
FT                   almost 2-fold increase in affinity."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   MUTAGEN         267
FT                   /note="H->A: Weakly active, 1000-fold decrease in catalytic
FT                   efficiency. Very slow ketonisation and C-C cleavage."
FT                   /evidence="ECO:0000269|PubMed:15663941"
FT   MUTAGEN         268
FT                   /note="W->G: 10-fold and 20-fold decrease in catalytic
FT                   activity and affinity, respectively."
FT                   /evidence="ECO:0000269|PubMed:17029402"
FT   CONFLICT        153
FT                   /note="E -> G (in Ref. 1; BAA13054)"
FT                   /evidence="ECO:0000305"
FT   HELIX           8..11
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          12..19
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          22..31
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          35..41
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           50..53
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   TURN            54..57
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           58..63
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          67..71
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           88..102
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          108..113
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           115..126
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           128..130
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          131..138
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          146..148
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           153..163
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           167..175
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           186..198
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           200..212
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           221..226
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          231..236
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          240..242
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           245..253
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   STRAND          258..264
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           269..272
FT                   /evidence="ECO:0007829|PDB:1U2E"
FT   HELIX           274..285
FT                   /evidence="ECO:0007829|PDB:1U2E"
SQ   SEQUENCE   288 AA;  31937 MW;  8F7E5ADE584A45BA CRC64;
     MSYQPQTEAA TSRFLNVEEA GKTLRIHFND CGQGDETVVL LHGSGPGATG WANFSRNIDP
     LVEAGYRVIL LDCPGWGKSD SVVNSGSRSD LNARILKSVV DQLDIAKIHL LGNSMGGHSS
     VAFTLKWPER VGKLVLMGGG TGGMSLFTPM PTEGIKRLNQ LYRQPTIENL KLMMDIFVFD
     TSDLTDALFE ARLNNMLSRR DHLENFVKSL EANPKQFPDF GPRLAEIKAQ TLIVWGRNDR
     FVPMDAGLRL LSGIAGSELH IFRDCGHWAQ WEHADAFNQL VLNFLARP
//