ID HMGA_PSEOC Reviewed; 227 AA. AC Q9AQI0; DT 08-FEB-2011, integrated into UniProtKB/Swiss-Prot. DT 01-JUN-2001, sequence version 1. DT 13-SEP-2023, entry version 64. DE RecName: Full=4-hydroxy-4-methyl-2-oxoglutarate aldolase/4-carboxy-4-hydroxy-2-oxoadipate aldolase; DE Short=HMG/CHA aldolase; DE EC=4.1.3.16 {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; DE EC=4.1.3.17 {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; DE AltName: Full=4-hydroxy-2-oxoglutarate aldolase; DE AltName: Full=Oxaloacetate decarboxylase; DE Short=OAA decarboxylase; DE EC=4.1.1.112 {ECO:0000269|PubMed:2229032}; GN Name=proA {ECO:0000312|EMBL:BAB21456.3}; OS Pseudomonas straminea. OC Bacteria; Pseudomonadota; Gammaproteobacteria; Pseudomonadales; OC Pseudomonadaceae; Pseudomonas. OX NCBI_TaxID=47882; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-22, FUNCTION, RP CATALYTIC ACTIVITY, COFACTOR, AND BIOPHYSICOCHEMICAL PROPERTIES. RC STRAIN=NGJ1 {ECO:0000312|EMBL:BAB21456.3}; RX PubMed=11826967; DOI=10.1271/bbb.65.2701; RA Maruyama K., Miwa M., Tsujii N., Nagai T., Tomita N., Harada T., RA Sobajima H., Sugisaki H.; RT "Cloning, sequencing, and expression of the gene encoding 4-hydroxy-4- RT methyl-2-oxoglutarate aldolase from Pseudomonas ochraceae NGJ1."; RL Biosci. Biotechnol. Biochem. 65:2701-2709(2001). RN [2] RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND INDUCTION. RX PubMed=2229032; DOI=10.1093/oxfordjournals.jbchem.a123201; RA Maruyama K.; RT "Purification and properties of 4-hydroxy-4-methyl-2-oxoglutarate aldolase RT from Pseudomonas ochraceae grown on phthalate."; RL J. Biochem. 108:327-333(1990). RN [3] RP ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES. RX PubMed=2229033; DOI=10.1093/oxfordjournals.jbchem.a123202; RA Maruyama K.; RT "Activation of Pseudomonas ochraceae 4-hydroxy-4-methyl-2-oxoglutarate RT aldolase by inorganic phosphate."; RL J. Biochem. 108:334-340(1990). RN [4] RP ACTIVITY REGULATION. RX PubMed=1794988; DOI=10.1093/oxfordjournals.jbchem.a123699; RA Maruyama K.; RT "Chemical modification of Pseudomonas ochraceae 4-hydroxy-4-methyl-2- RT oxoglutarate aldolase by diethyl pyrocarbonate."; RL J. Biochem. 110:976-981(1991). CC -!- FUNCTION: Catalyzes the last step of the bacterial protocatechuate 4,5- CC cleavage pathway. Has a broad substrate specificity and catalyzes the CC aldol cleavage of 4-hydroxy-4-methyl-2-oxoglutarate, 4-hydroxy-2- CC oxoglutarate and 4-carboxy-4-hydroxy-2-oxoadipate, and the CC decarboxylation of oxaloacetate. Preferentially cleaves the L-isomer of CC 4-carboxy-4-hydroxy-2-oxoadipate, and has lower activity towards 4- CC hydroxy-4-methyl-2-oxoglutarate and 4-Hydroxy-2-oxoglutarate. Does not CC cleave 4-hydroxy-2-oxovalerate, citrate, 4-hydroxy-2-oxobutyrate, 2- CC oxoglutarate or fructose-1,6-bisphosphate. CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}. CC -!- CATALYTIC ACTIVITY: CC Reaction=(4S)-4-hydroxy-2-oxoglutarate = glyoxylate + pyruvate; CC Xref=Rhea:RHEA:35639, ChEBI:CHEBI:15361, ChEBI:CHEBI:36655, CC ChEBI:CHEBI:71685; EC=4.1.3.16; CC Evidence={ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; CC -!- CATALYTIC ACTIVITY: CC Reaction=(4R)-4-hydroxy-2-oxoglutarate = glyoxylate + pyruvate; CC Xref=Rhea:RHEA:30687, ChEBI:CHEBI:15361, ChEBI:CHEBI:36655, CC ChEBI:CHEBI:62213; EC=4.1.3.16; CC Evidence={ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; CC -!- CATALYTIC ACTIVITY: CC Reaction=4-hydroxy-4-methyl-2-oxoglutarate = 2 pyruvate; CC Xref=Rhea:RHEA:22748, ChEBI:CHEBI:15361, ChEBI:CHEBI:58276; CC EC=4.1.3.17; Evidence={ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032}; CC -!- CATALYTIC ACTIVITY: CC Reaction=2-hydroxy-4-oxobutane-1,2,4-tricarboxylate = oxaloacetate + CC pyruvate; Xref=Rhea:RHEA:28935, ChEBI:CHEBI:15361, ChEBI:CHEBI:16452, CC ChEBI:CHEBI:58075; EC=4.1.3.17; CC Evidence={ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; CC -!- CATALYTIC ACTIVITY: CC Reaction=H(+) + oxaloacetate = CO2 + pyruvate; Xref=Rhea:RHEA:15641, CC ChEBI:CHEBI:15361, ChEBI:CHEBI:15378, ChEBI:CHEBI:16452, CC ChEBI:CHEBI:16526; EC=4.1.1.112; CC Evidence={ECO:0000269|PubMed:2229032}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; CC Note=Divalent metal cations. Probably Mg(2+). CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032}; CC -!- ACTIVITY REGULATION: Cleavage of 4-carboxy-4-hydroxy-2-oxoadipate, and CC to a lesser extent 4-hydroxy-4-methyl-2-oxoglutarate, is inhibited by CC lysine modification caused by diethyl pyrocarbonate. Decarboxylation of CC oxaloacetate is unaffected by diethyl pyrocarbonate. Inhibited by CC BeCl(2), CaCl(2), NiCl(2), BaCl(2), HgCl(2), SrSO(4), CrCl(3) and CC FeCl(3). Partially inhibited by p-chloromercuribenzoate and N- CC ethylmaleimide. Activated by inorganic phosphate, arsenate, phosphorous CC acid, acetyl phosphate, thiamine diphosphate, ADP, ATP and diphosphate. CC {ECO:0000269|PubMed:1794988, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.044 mM for DL-4-carboxy-4-hydroxy-2-oxoadipate CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC KM=0.019 mM for L-4-carboxy-4-hydroxy-2-oxoadipate CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC KM=0.15 mM for D-4-carboxy-4-hydroxy-2-oxoadipate CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC KM=1.25 mM for DL-4-hydroxy-4-methyl-2-oxoglutarate CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC KM=0.24 mM for DL-4-hydroxy-2-oxoglutarate CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC KM=0.5 mM for oxaloacetate {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC Vmax=1250 umol/min/mg enzyme with DL-4-carboxy-4-hydroxy-2-oxoadipate CC as substrate {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC Vmax=1220 umol/min/mg enzyme with L-4-carboxy-4-hydroxy-2-oxoadipate CC as substrate {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC Vmax=67.6 umol/min/mg enzyme with D-4-carboxy-4-hydroxy-2-oxoadipate CC as substrate {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC Vmax=213 umol/min/mg enzyme with DL-4-hydroxy-4-methyl-2-oxoglutarate CC as substrate {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC Vmax=1.3 umol/min/mg enzyme with DL-4-hydroxy-2-oxoglutarate as CC substrate {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC Vmax=20.8 umol/min/mg enzyme with oxaloacetate as substrate CC {ECO:0000269|PubMed:11826967, ECO:0000269|PubMed:2229032, CC ECO:0000269|PubMed:2229033}; CC pH dependence: CC Optimum pH varies depending on the substrate used and phosphate CC concentration. In the absence of inorganic phosphate pH optima are CC 6.6 for L-4-carboxy-4-hydroxy-2-oxoadipate, 8.0 for CC D-4-carboxy-4-hydroxy-2-oxoadipate, 6.7 and 8.0 for CC DL-4-carboxy-4-hydroxy-2-oxoadipate, 8.3 for CC DL-4-hydroxy-4-methyl-2-oxoglutarate, 9.3 for CC DL-4-hydroxy-2-oxoglutarate and 8.8 for oxaloacetate. In the presence CC of 3 mM inorganic phosphate pH optima are more alkaline: 8.2 for CC L-4-carboxy-4-hydroxy-2-oxoadipate, 8.6 for CC D-4-carboxy-4-hydroxy-2-oxoadipate, 8.2 for CC DL-4-carboxy-4-hydroxy-2-oxoadipate, 8.9 for CC DL-4-hydroxy-4-methyl-2-oxoglutarate, 9.4 for CC DL-4-hydroxy-2-oxoglutarate and 8.9 for oxaloacetate. Stable at pH CC 6.0 to pH 9.5. {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC Temperature dependence: CC Retains 50% of maximum activity after incubation at 54 degrees CC Celsius for 10 minutes. {ECO:0000269|PubMed:11826967, CC ECO:0000269|PubMed:2229032, ECO:0000269|PubMed:2229033}; CC -!- SUBUNIT: Homohexamer. {ECO:0000269|PubMed:2229032}. CC -!- INDUCTION: By growth on aromatic carboxylates such as phthalate, CC terephthalate, m-hydroxybenzoate and p-hydroxybenzoate. CC {ECO:0000269|PubMed:2229032}. CC -!- SIMILARITY: Belongs to the LigK/PcmE family. {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AB050935; BAB21456.3; -; Genomic_DNA. DR AlphaFoldDB; Q9AQI0; -. DR SMR; Q9AQI0; -. DR BioCyc; MetaCyc:MONOMER-3244; -. DR GO; GO:0106009; F:(4S)-4-hydroxy-2-oxoglutarate aldolase activity; IEA:RHEA. DR GO; GO:0008700; F:4-hydroxy-2-oxoglutarate aldolase activity; IDA:UniProtKB. DR GO; GO:0047443; F:4-hydroxy-4-methyl-2-oxoglutarate aldolase activity; IDA:UniProtKB. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0008948; F:oxaloacetate decarboxylase activity; IDA:UniProtKB. DR GO; GO:0019619; P:3,4-dihydroxybenzoate catabolic process; IDA:UniProtKB. DR CDD; cd16841; RraA_family; 1. DR Gene3D; 3.50.30.40; Ribonuclease E inhibitor RraA/RraA-like; 1. DR InterPro; IPR014165; LigK_PcmE. DR InterPro; IPR005493; RraA/RraA-like. DR InterPro; IPR036704; RraA/RraA-like_sf. DR NCBIfam; TIGR02798; ligK_PcmE; 1. DR PANTHER; PTHR33254; 4-HYDROXY-4-METHYL-2-OXOGLUTARATE ALDOLASE 3-RELATED; 1. DR PANTHER; PTHR33254:SF16; BLR3842 PROTEIN; 1. DR Pfam; PF03737; RraA-like; 1. DR SUPFAM; SSF89562; RraA-like; 1. PE 1: Evidence at protein level; KW Direct protein sequencing; Lyase; Magnesium; Metal-binding. FT CHAIN 1..227 FT /note="4-hydroxy-4-methyl-2-oxoglutarate aldolase/4- FT carboxy-4-hydroxy-2-oxoadipate aldolase" FT /id="PRO_0000403973" FT BINDING 97..100 FT /ligand="substrate" FT /evidence="ECO:0000250|UniProtKB:A5W059" FT BINDING 119 FT /ligand="substrate" FT /evidence="ECO:0000250|UniProtKB:A5W059" FT BINDING 120 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT /evidence="ECO:0000250|UniProtKB:A5W059" SQ SEQUENCE 227 AA; 24068 MW; F56501D5BDD0262F CRC64; MYELGVVYRN IQRADRAAAD GLAALGSATV HEAMGRVGLL KPYMRPIYAG KQVSGTAVTV LLQPGDNWMM HVAAEQIQPG DIVVAAVTAE CTDGYFGDLL ATSFQARGAR ALIIDAGVRD VKTLQEMDFP VWSKAISSKG TIKATLGSVN IPIVCAGMLV TPGDVIVADD DGVVCVPAAR AVEVLAAAQK RESFEGEKRA KLASGVLGLD MYKMREPLEK AGLKYID //