HOA_THET8
ID HOA_THET8 Reviewed; 347 AA.
AC Q53WI0;
DT 03-NOV-2009, integrated into UniProtKB/Swiss-Prot.
DT 24-MAY-2005, sequence version 1.
DT 03-AUG-2022, entry version 87.
DE RecName: Full=4-hydroxy-2-oxovalerate aldolase {ECO:0000255|HAMAP-Rule:MF_01656};
DE Short=HOA {ECO:0000255|HAMAP-Rule:MF_01656};
DE EC=4.1.3.39 {ECO:0000255|HAMAP-Rule:MF_01656, ECO:0000269|PubMed:22316175};
DE AltName: Full=4-hydroxy-2-keto-pentanoic acid aldolase {ECO:0000255|HAMAP-Rule:MF_01656};
DE AltName: Full=4-hydroxy-2-oxohexanoate aldolase;
DE EC=4.1.3.43 {ECO:0000269|PubMed:22316175};
DE AltName: Full=4-hydroxy-2-oxopentanoate aldolase {ECO:0000255|HAMAP-Rule:MF_01656};
GN OrderedLocusNames=TTHB246;
OS Thermus thermophilus (strain ATCC 27634 / DSM 579 / HB8).
OG Plasmid pTT27.
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=300852;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 27634 / DSM 579 / HB8;
RA Masui R., Kurokawa K., Nakagawa N., Tokunaga F., Koyama Y., Shibata T.,
RA Oshima T., Yokoyama S., Yasunaga T., Kuramitsu S.;
RT "Complete genome sequence of Thermus thermophilus HB8.";
RL Submitted (NOV-2004) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBSTRATE SPECIFICITY, ACTIVITY REGULATION, SUBUNIT, COMPLEX WITH TTHB247,
RP ALDEHYDE CHANNELING, AND MUTAGENESIS OF ALA-324.
RC STRAIN=ATCC 27634 / DSM 579 / HB8;
RX PubMed=22316175; DOI=10.1021/bi201832a;
RA Baker P., Hillis C., Carere J., Seah S.Y.;
RT "Protein-protein interactions and substrate channeling in orthologous and
RT chimeric aldolase-dehydrogenase complexes.";
RL Biochemistry 51:1942-1952(2012).
CC -!- FUNCTION: Catalyzes the retro-aldol cleavage of both 4-hydroxy-2-
CC oxopentanoate (HOPA) and 4-hydroxy-2-oxohexanoate (HOHA) to pyruvate
CC and acetaldehyde or propanaldehyde, respectively. The aldehydes
CC produced by this reaction are directly channeled to the dehydrogenase
CC TTHB247, ensuring that these toxic aldehydes are sequestered from
CC cellular components. Is involved in the meta-cleavage pathway for the
CC degradation of aromatic compounds. Appears to be stereospecific since
CC it can cleave (4S)-4-hydroxy-2-oxopentanoate but not the (4R) isomer.
CC Is not able to catalyze the aldol addition of 2-oxobutyrate with
CC acetaldehyde; this indicates that the enzyme is specific for pyruvate
CC as the carbonyl donor. {ECO:0000269|PubMed:22316175}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-4-hydroxy-2-oxopentanoate = acetaldehyde + pyruvate;
CC Xref=Rhea:RHEA:22624, ChEBI:CHEBI:15343, ChEBI:CHEBI:15361,
CC ChEBI:CHEBI:73143; EC=4.1.3.39; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_01656, ECO:0000269|PubMed:22316175};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-4-hydroxy-2-oxohexanoate = propanal + pyruvate;
CC Xref=Rhea:RHEA:36003, ChEBI:CHEBI:15361, ChEBI:CHEBI:17153,
CC ChEBI:CHEBI:73142; EC=4.1.3.43;
CC Evidence={ECO:0000269|PubMed:22316175};
CC -!- COFACTOR:
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:22316175};
CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC Evidence={ECO:0000269|PubMed:22316175};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:22316175};
CC Note=Divalent metal cation. Has the highest activity with Co(2+) as
CC cofactor, followed by Ni(2+) and Mn(2+). Mg(2+) and Ca(2+) are very
CC poor metal cofactors. {ECO:0000269|PubMed:22316175};
CC -!- ACTIVITY REGULATION: Appears to be allosterically activated by NADH.
CC {ECO:0000269|PubMed:22316175}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=206 uM for 4-hydroxy-2-oxopentanoate (in the presence of NADH at
CC pH 8 and 25 degrees Celsius) {ECO:0000269|PubMed:22316175};
CC KM=41 uM for 4-hydroxy-2-oxopentanoate (in the presence of NADH and
CC in complex with the dehydrogenase TTHB247, at pH 8 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:22316175};
CC KM=123 uM for 4-hydroxy-2-oxopentanoate (in the absence of NADH and
CC in complex with the dehydrogenase TTHB247, at pH 8 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:22316175};
CC KM=210 uM for 4-hydroxy-2-oxohexanoate (in the presence of NADH at pH
CC 8 and 25 degrees Celsius) {ECO:0000269|PubMed:22316175};
CC KM=37 uM for 4-hydroxy-2-oxopentanoate (in the presence of NADH and
CC in complex with the dehydrogenase TTHB247, at pH 8 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:22316175};
CC Note=The catalytic efficiency is similar when using 4-hydroxy-2-
CC oxopentanoate or 4-hydroxy-2-oxohexanoate as substrate. In the
CC presence of NADH, exhibits a 2-fold increase in kcat.;
CC Temperature dependence:
CC Has a half-life of 42 hours at 50 degrees Celsius. When TTHB246 is in
CC complex with TTHB247, its half-life is reduced by approximately 30
CC hours. {ECO:0000269|PubMed:22316175};
CC -!- SUBUNIT: Homodimer. Can also form a heterotetramer composed of two
CC aldolase (TTHB246) and two dehydrogenase (TTHB247) subunits. Upon
CC complex formation, the aldolase shows a 5-fold increase in substrate
CC affinity, while the dehydrogenase shows a 3-fold decrease; the kcat
CC values of each enzyme are reduced by 2-fold when they are in a complex.
CC {ECO:0000269|PubMed:22316175}.
CC -!- SIMILARITY: Belongs to the 4-hydroxy-2-oxovalerate aldolase family.
CC {ECO:0000255|HAMAP-Rule:MF_01656, ECO:0000305}.
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DR EMBL; AP008227; BAD72042.1; -; Genomic_DNA.
DR RefSeq; WP_011229063.1; NC_006462.1.
DR RefSeq; YP_145485.1; NC_006462.1.
DR AlphaFoldDB; Q53WI0; -.
DR SMR; Q53WI0; -.
DR EnsemblBacteria; BAD72042; BAD72042; BAD72042.
DR GeneID; 3169557; -.
DR KEGG; ttj:TTHB246; -.
DR PATRIC; fig|300852.9.peg.2202; -.
DR HOGENOM; CLU_049173_0_0_0; -.
DR OMA; DLYKMMD; -.
DR PhylomeDB; Q53WI0; -.
DR BRENDA; 4.1.3.43; 2305.
DR SABIO-RK; Q53WI0; -.
DR Proteomes; UP000000532; Plasmid pTT27.
DR GO; GO:0008701; F:4-hydroxy-2-oxovalerate aldolase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0030145; F:manganese ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-UniRule.
DR CDD; cd07943; DRE_TIM_HOA; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR HAMAP; MF_01656; HOA; 1.
DR InterPro; IPR017629; 4OH_2_O-val_aldolase.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR012425; DmpG_comm.
DR InterPro; IPR035685; DRE_TIM_HOA.
DR InterPro; IPR000891; PYR_CT.
DR Pfam; PF07836; DmpG_comm; 1.
DR Pfam; PF00682; HMGL-like; 1.
DR TIGRFAMs; TIGR03217; 4OH_2_O_val_ald; 1.
DR PROSITE; PS50991; PYR_CT; 1.
PE 1: Evidence at protein level;
KW Allosteric enzyme; Aromatic hydrocarbons catabolism; Cobalt; Lyase;
KW Manganese; Metal-binding; Nickel; Plasmid; Reference proteome.
FT CHAIN 1..347
FT /note="4-hydroxy-2-oxovalerate aldolase"
FT /id="PRO_0000387928"
FT DOMAIN 11..262
FT /note="Pyruvate carboxyltransferase"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT ACT_SITE 23
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 19..20
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 20
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 173
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 201
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 201
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 203
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT BINDING 292
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT SITE 19
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01656"
FT MUTAGEN 324
FT /note="A->G: Increases the channeling efficiency of
FT propanaldehyde from 57% to 94%."
FT /evidence="ECO:0000269|PubMed:22316175"
SQ SEQUENCE 347 AA; 37214 MW; 3E9DC806D6EAC64D CRC64;
MSWDLSTAKP PVVVDTTLRD GSHAHRHQYT VEEARAIAQA LDEAGVYAIE VSHGDGLGGS
SLQYGFSRTD EMELIRAVRE TVRRAKVAAL LLPGIGTRKE LKEAVEAGIQ MVRIATQCTE
ADISEQHFGM AKEMGLEAVG FLMMSHMRPP EFLAEQARLM EGYGADVVYI VDSAGAMLPE
DAYARVKALK EALSRAKVGF HAHNNLGLAI GNTLAALAAG ADWVDATLRG YGAGAGNAPL
EVLAAVLDKA GLNPGLDVFK LLDAAEYVMG PILHFQPYPD RDSVAIGYAG VYSTFLLHAK
RIGKELGVDP LAILLELGRR QAVAGQEDWI LRVALELKEK EAGALAD
