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 2-hydroxy-6-
CC oxononadienedioate as substrate. {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
//
