HPCH_ECOLX
ID HPCH_ECOLX Reviewed; 262 AA.
AC Q47098; Q46983;
DT 01-DEC-2000, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 116.
DE RecName: Full=4-hydroxy-2-oxo-heptane-1,7-dioate aldolase;
DE EC=4.1.2.52;
DE AltName: Full=2,4-dihydroxyhept-2-ene-1,7-dioic acid aldolase;
DE Short=HHED aldolase;
DE AltName: Full=4-hydroxy-2-ketoheptane-1,7-dioate aldolase;
DE Short=HKHD aldolase;
GN Name=hpcH; Synonyms=hpaI;
OS Escherichia coli.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 1-10.
RC STRAIN=C;
RX PubMed=8529896; DOI=10.1016/0378-1119(95)00596-8;
RA Stringfellow J.M., Turpin B., Cooper R.A.;
RT "Sequence of the Escherichia coli C homoprotocatechuic acid degradative
RT operon completed with that of the 2,4-dihydroxyhept-2-ene-1,7-dioic acid
RT aldolase-encoding gene (hpcH).";
RL Gene 166:73-76(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=W / ATCC 11105 / DSM 1900 / 113-3;
RX PubMed=8550403; DOI=10.1128/jb.178.1.111-120.1996;
RA Prieto M.A., Diaz E., Garcia J.L.;
RT "Molecular characterization of the 4-hydroxyphenylacetate catabolic pathway
RT of Escherichia coli W: engineering a mobile aromatic degradative cluster.";
RL J. Bacteriol. 178:111-120(1996).
RN [3]
RP CRYSTALLIZATION.
RC STRAIN=C;
RX PubMed=16511168; DOI=10.1107/s1744309105023079;
RA Rea D., Fueloep V., Bugg T.D.H., Roper D.I.;
RT "Expression, purification and preliminary crystallographic analysis of 2,4-
RT dihydroxy-hepta-2-ene-1,7-dioate aldolase (HpcH) from Escherichia coli C.";
RL Acta Crystallogr. F 61:821-824(2005).
RN [4]
RP FUNCTION, COFACTOR, SUBSTRATE SPECIFICITY, KINETIC PARAMETERS, ACTIVITY
RP REGULATION, AND MUTAGENESIS OF ARG-70.
RX PubMed=15996099; DOI=10.1021/bi050607y;
RA Wang W., Seah S.Y.K.;
RT "Purification and biochemical characterization of a pyruvate-specific class
RT II aldolase, HpaI.";
RL Biochemistry 44:9447-9455(2005).
RN [5]
RP MUTAGENESIS OF HIS-45, AND ACTIVE SITE.
RX PubMed=18775708; DOI=10.1016/j.febslet.2008.08.032;
RA Wang W., Seah S.Y.K.;
RT "The role of a conserved histidine residue in a pyruvate-specific Class II
RT aldolase.";
RL FEBS Lett. 582:3385-3388(2008).
RN [6]
RP FUNCTION, SUBSTRATE SPECIFICITY, STEREOSPECIFICITY, KINETIC PARAMETERS, AND
RP KINETIC MECHANISM.
RX PubMed=20364820; DOI=10.1021/bi100251u;
RA Wang W., Baker P., Seah S.Y.;
RT "Comparison of two metal-dependent pyruvate aldolases related by convergent
RT evolution: substrate specificity, kinetic mechanism, and substrate
RT channeling.";
RL Biochemistry 49:3774-3782(2010).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH
RP MAGNESIUM AND SUBSTRATE ANALOG, SUBUNIT, CATALYTIC MECHANISM, AND
RP MUTAGENESIS OF HIS-45.
RC STRAIN=C;
RX PubMed=17881002; DOI=10.1016/j.jmb.2007.06.048;
RA Rea D., Fueloep V., Bugg T.D.H., Roper D.I.;
RT "Structure and mechanism of HpcH: a metal ion dependent class II aldolase
RT from the homoprotocatechuate degradation pathway of Escherichia coli.";
RL J. Mol. Biol. 373:866-876(2007).
CC -!- FUNCTION: Catalyzes the reversible retro-aldol cleavage of 4-hydroxy-2-
CC ketoheptane-1,7-dioate (HKHD) to pyruvate and succinate semialdehyde.
CC Is also able to catalyze the aldol cleavage of 4-hydroxy-2-
CC ketopentanoate and 4-hydroxy-2-ketohexanoate. Is not stereospecific
CC since it can cleave both substrate enantiomers. Also exhibits
CC significant oxaloacetate decarboxylase activity in vitro. In the
CC reverse direction, is able to condense a range of aldehyde acceptors
CC (from two to five carbons in length) with pyruvate or 2-oxobutanoate.
CC Unlike with BphI from Burkholderia xenovorans, the aldol addition
CC reaction lacks stereospecificity, producing a racemic mixture.
CC {ECO:0000269|PubMed:15996099, ECO:0000269|PubMed:20364820}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-hydroxy-2-oxoheptanedioate = pyruvate + succinate
CC semialdehyde; Xref=Rhea:RHEA:25788, ChEBI:CHEBI:15361,
CC ChEBI:CHEBI:57706, ChEBI:CHEBI:73036; EC=4.1.2.52;
CC -!- COFACTOR:
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:15996099};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:15996099};
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:15996099};
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:15996099};
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:15996099};
CC Note=Binds 1 divalent metal cation per subunit. Has the highest
CC activity with Co(2+) and Mn(2+), but can also use Zn(2+), Fe(2+) and
CC Mg(2+). {ECO:0000269|PubMed:15996099};
CC -!- ACTIVITY REGULATION: Competitively inhibited by oxalate.
CC {ECO:0000269|PubMed:15996099}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.38 mM for 4-hydroxy-2-ketopentanoate
CC {ECO:0000269|PubMed:15996099};
CC KM=0.16 mM for 4-hydroxy-2-ketohexanoate
CC {ECO:0000269|PubMed:15996099};
CC KM=14.22 mM for 3-deoxy-D-manno-oct-2-ulosonate (KDO)
CC {ECO:0000269|PubMed:15996099};
CC KM=1.37 mM for oxaloacetate {ECO:0000269|PubMed:15996099,
CC ECO:0000269|PubMed:20364820};
CC KM=9.1 mM for succinate semialdehyde {ECO:0000269|PubMed:20364820};
CC KM=62.9 mM for acetaldehyde {ECO:0000269|PubMed:20364820};
CC KM=33.3 mM for glycolaldehyde {ECO:0000269|PubMed:20364820};
CC KM=32.9 mM for propanaldehyde {ECO:0000269|PubMed:20364820};
CC KM=6.0 uM for Co(2+) {ECO:0000269|PubMed:15996099};
CC KM=17.5 uM for Mn(2+) {ECO:0000269|PubMed:15996099};
CC KM=289.3 uM for Mg(2+) {ECO:0000269|PubMed:15996099};
CC Note=The catalytic efficiency measured with 3-deoxy-D-manno-oct-2-
CC ulosonate (KDO) is 37800-fold lower than that with 4-hydroxy-2-
CC ketohexanoate. {ECO:0000269|PubMed:15996099};
CC -!- PATHWAY: Aromatic compound metabolism; 4-hydroxyphenylacetate
CC degradation; pyruvate and succinate semialdehyde from 4-
CC hydroxyphenylacetate: step 7/7.
CC -!- SUBUNIT: Homohexamer; trimer of dimers. {ECO:0000269|PubMed:17881002}.
CC -!- MISCELLANEOUS: The aldol addition reaction proceeds via a rapid
CC equilibrium random order mechanism.
CC -!- SIMILARITY: Belongs to the HpcH/HpaI aldolase family. {ECO:0000305}.
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DR EMBL; Z47799; CAA87759.1; -; Genomic_DNA.
DR EMBL; Z37980; CAA86045.1; -; Genomic_DNA.
DR PIR; S49311; S49311.
DR RefSeq; WP_000431706.1; NZ_WVVQ01000010.1.
DR PDB; 2V5J; X-ray; 1.60 A; A/B=1-262.
DR PDB; 2V5K; X-ray; 2.20 A; A/B=1-262.
DR PDBsum; 2V5J; -.
DR PDBsum; 2V5K; -.
DR AlphaFoldDB; Q47098; -.
DR SMR; Q47098; -.
DR STRING; 585034.ECIAI1_4570; -.
DR eggNOG; COG3836; Bacteria.
DR OMA; WNRVDDY; -.
DR OrthoDB; 1797448at2; -.
DR BioCyc; MetaCyc:MON-2849; -.
DR BRENDA; 4.1.2.52; 2026.
DR BRENDA; 4.1.3.39; 2026.
DR SABIO-RK; Q47098; -.
DR UniPathway; UPA00208; UER00422.
DR EvolutionaryTrace; Q47098; -.
DR GO; GO:0018802; F:2,4-dihydroxyhept-2-ene-1,7-dioate aldolase activity; IEA:InterPro.
DR GO; GO:0043863; F:4-hydroxy-2-ketopimelate aldolase activity; IEA:RHEA.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0010124; P:phenylacetate catabolic process; IEA:InterPro.
DR Gene3D; 3.20.20.60; -; 1.
DR HAMAP; MF_01292; HKHD_aldolase; 1.
DR InterPro; IPR005000; Aldolase/citrate-lyase_domain.
DR InterPro; IPR023701; HKHD_aldolase_ent.
DR InterPro; IPR012689; HpaI.
DR InterPro; IPR015813; Pyrv/PenolPyrv_Kinase-like_dom.
DR InterPro; IPR040442; Pyrv_Kinase-like_dom_sf.
DR Pfam; PF03328; HpcH_HpaI; 1.
DR SUPFAM; SSF51621; SSF51621; 1.
DR TIGRFAMs; TIGR02311; HpaI; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aromatic hydrocarbons catabolism; Cobalt;
KW Direct protein sequencing; Iron; Lyase; Magnesium; Manganese;
KW Metal-binding; Zinc.
FT CHAIN 1..262
FT /note="4-hydroxy-2-oxo-heptane-1,7-dioate aldolase"
FT /id="PRO_0000207095"
FT ACT_SITE 45
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:17881002,
FT ECO:0000269|PubMed:18775708"
FT BINDING 147
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:17881002"
FT BINDING 149
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /evidence="ECO:0000269|PubMed:17881002"
FT BINDING 174
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:17881002"
FT BINDING 175
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /evidence="ECO:0000269|PubMed:17881002"
FT BINDING 175
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:17881002"
FT SITE 70
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:17881002"
FT SITE 84
FT /note="Increases basicity of active site His"
FT /evidence="ECO:0000305|PubMed:17881002"
FT MUTAGEN 45
FT /note="H->A,Q: Loss of activity."
FT /evidence="ECO:0000269|PubMed:17881002,
FT ECO:0000269|PubMed:18775708"
FT MUTAGEN 70
FT /note="R->A: Loss of activity. Still able to bind
FT pyruvate."
FT /evidence="ECO:0000269|PubMed:15996099"
FT HELIX 4..10
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 15..20
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 25..32
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 37..47
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 50..60
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 63..70
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 72..74
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 76..84
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 89..93
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 98..107
FT /evidence="ECO:0007829|PDB:2V5J"
FT TURN 111..113
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 118..120
FT /evidence="ECO:0007829|PDB:2V5J"
FT TURN 121..123
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 124..126
FT /evidence="ECO:0007829|PDB:2V5J"
FT TURN 127..130
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 134..141
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 143..148
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 151..155
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 157..161
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 166..171
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 173..179
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 189..204
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 207..212
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 216..224
FT /evidence="ECO:0007829|PDB:2V5J"
FT STRAND 228..234
FT /evidence="ECO:0007829|PDB:2V5J"
FT HELIX 235..257
FT /evidence="ECO:0007829|PDB:2V5J"
SQ SEQUENCE 262 AA; 28073 MW; 374F576B6C53587F CRC64;
MENSFKAALK AGRPQIGLWL GLSSSYSAEL LAGAGFDWLL IDGEHAPNNV QTVLTQLQAI
APYPSQPVVR PSWNDPVQIK QLLDVGTQTL LVPMVQNADE AREAVRATRY PPAGIRGVGS
ALARASRWNR IPDYLQKAND QMCVLVQIET REAMKNLPQI LDVEGVDGVF IGPADLSADM
GYAGNPQHPE VQAAIEQAIV QIRESGKAPG ILIANEQLAK RYLELGALFV AVGVDTTLLA
RAAEALAARF GAQATAVKPG VY
