HAOX2_RAT
ID HAOX2_RAT Reviewed; 353 AA.
AC Q07523;
DT 01-OCT-1996, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 2.
DT 03-AUG-2022, entry version 156.
DE RecName: Full=2-Hydroxyacid oxidase 2;
DE Short=HAOX2;
DE EC=1.1.3.15 {ECO:0000305|PubMed:15683236, ECO:0000305|PubMed:3061453, ECO:0000305|PubMed:8508789};
DE AltName: Full=(S)-2-hydroxy-acid oxidase, peroxisomal;
DE AltName: Full=Long chain alpha-hydroxy acid oxidase {ECO:0000303|PubMed:1939137};
DE AltName: Full=Long-chain L-2-hydroxy acid oxidase {ECO:0000303|PubMed:15683236, ECO:0000303|PubMed:8508789};
DE Short=LCHAO {ECO:0000303|PubMed:15683236};
GN Name=Hao2; Synonyms=Hao3, Haox2;
OS Rattus norvegicus (Rat).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC Murinae; Rattus.
OX NCBI_TaxID=10116;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], TISSUE SPECIFICITY, SUBUNIT, COFACTOR,
RP CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=Wistar; TISSUE=Kidney;
RX PubMed=8508789; DOI=10.1111/j.1432-1033.1993.tb17891.x;
RA Belmouden A., Le K.H.D., Lederer F., Garchon H.J.;
RT "Molecular cloning and nucleotide sequence of cDNA encoding rat kidney
RT long-chain L-2-hydroxy acid oxidase. Expression of the catalytically active
RT recombinant protein as a chimaera.";
RL Eur. J. Biochem. 214:17-25(1993).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC TISSUE=Testis;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA project:
RT the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [3]
RP PROTEIN SEQUENCE OF 2-353.
RC TISSUE=Kidney;
RX PubMed=1939137; DOI=10.1016/s0021-9258(18)54791-8;
RA Le K.H.D., Lederer F.;
RT "Amino acid sequence of long chain alpha-hydroxy acid oxidase from rat
RT kidney, a member of the family of FMN-dependent alpha-hydroxy acid-
RT oxidizing enzymes.";
RL J. Biol. Chem. 266:20877-20881(1991).
RN [4]
RP PROTEIN SEQUENCE OF 2-41, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND COFACTOR.
RX PubMed=3061453; DOI=10.1021/bi00419a029;
RA Urban P., Chirat I., Lederer F.;
RT "Rat kidney L-2-hydroxyacid oxidase. Structural and mechanistic comparison
RT with flavocytochrome b2 from baker's yeast.";
RL Biochemistry 27:7365-7371(1988).
RN [5]
RP PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-133, AND IDENTIFICATION BY
RP MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX PubMed=22673903; DOI=10.1038/ncomms1871;
RA Lundby A., Secher A., Lage K., Nordsborg N.B., Dmytriyev A., Lundby C.,
RA Olsen J.V.;
RT "Quantitative maps of protein phosphorylation sites across 14 different rat
RT organs and tissues.";
RL Nat. Commun. 3:876-876(2012).
RN [6] {ECO:0007744|PDB:1TB3}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH FMN, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, SUBUNIT, AND
RP ACTIVE SITE.
RX PubMed=15683236; DOI=10.1021/bi048616e;
RA Cunane L.M., Barton J.D., Chen Z.-W., Le K.H.D., Amar D., Lederer F.,
RA Mathews F.S.;
RT "Crystal structure analysis of recombinant rat kidney long chain hydroxy
RT acid oxidase.";
RL Biochemistry 44:1521-1531(2005).
RN [7] {ECO:0007744|PDB:3SGZ}
RP X-RAY CRYSTALLOGRAPHY (1.35 ANGSTROMS) OF 2-353 IN COMPLEX WITH
RP 4-CARBOXY-5-[(4-CHLOROPHENYL)SULFANYL]-1,2,3-THIADIAZOLE INHIBITOR AND FMN,
RP COFACTOR, AND ACTIVITY REGULATION.
RX PubMed=22342614; DOI=10.1016/j.biochi.2012.02.003;
RA Chen Z.W., Vignaud C., Jaafar A., Levy B., Gueritte F., Guenard D.,
RA Lederer F., Mathews F.S.;
RT "High resolution crystal structure of rat long chain hydroxy acid oxidase
RT in complex with the inhibitor 4-carboxy-5-[(4-chlorophenyl)sulfanyl]-1,2,3-
RT thiadiazole. Implications for inhibitor specificity and drug design.";
RL Biochimie 94:1172-1179(2012).
CC -!- FUNCTION: Oxidase that catalyzes the oxidation of medium and long chain
CC hydroxyacids such as 2-hydroxyhexadecanoate, 2-hydroxyoctanoate, 2-
CC hydroxyhexanoate and 2-hydroxybutanoate, to the correspondong 2-
CC oxoacids (PubMed:15683236, PubMed:3061453, PubMed:8508789). Its role in
CC the oxidation of 2-hydroxy fatty acids may contribute to the general
CC pathway of fatty acid alpha-oxidation (By similarity). Can also use
CC mandelate as substrate (PubMed:3061453). Active in vitro with the
CC artificial electron acceptor 2,6-dichlorophenolindophenol (DCIP), but
CC O2 is believed to be the physiological electron acceptor, leading to
CC the production of H2O2 (PubMed:15683236, PubMed:3061453,
CC PubMed:8508789). {ECO:0000250|UniProtKB:Q9NYQ3,
CC ECO:0000269|PubMed:15683236, ECO:0000269|PubMed:3061453,
CC ECO:0000269|PubMed:8508789}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a (2S)-2-hydroxycarboxylate + O2 = a 2-oxocarboxylate + H2O2;
CC Xref=Rhea:RHEA:16789, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:35179, ChEBI:CHEBI:58123; EC=1.1.3.15;
CC Evidence={ECO:0000305|PubMed:15683236, ECO:0000305|PubMed:3061453,
CC ECO:0000305|PubMed:8508789};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:16790;
CC Evidence={ECO:0000305|PubMed:15683236, ECO:0000305|PubMed:3061453,
CC ECO:0000305|PubMed:8508789};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-hydroxyoctanoate + O2 = 2-oxooctanoate + H2O2;
CC Xref=Rhea:RHEA:67940, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:133514, ChEBI:CHEBI:176689;
CC Evidence={ECO:0000305|PubMed:15683236};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:67941;
CC Evidence={ECO:0000305|PubMed:15683236};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-hydroxyhexadecanoate + O2 = 2-oxohexadecanoate + H2O2;
CC Xref=Rhea:RHEA:67944, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:65097, ChEBI:CHEBI:176593;
CC Evidence={ECO:0000305|PubMed:15683236};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:67945;
CC Evidence={ECO:0000305|PubMed:15683236};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-hydroxyhexanoate + O2 = 2-oxohexanoate + H2O2;
CC Xref=Rhea:RHEA:69372, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:35177, ChEBI:CHEBI:133738;
CC Evidence={ECO:0000305|PubMed:3061453};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:69373;
CC Evidence={ECO:0000305|PubMed:3061453};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=mandelate + O2 = H2O2 + phenylglyoxylate;
CC Xref=Rhea:RHEA:68968, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:25147, ChEBI:CHEBI:36656;
CC Evidence={ECO:0000305|PubMed:3061453};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68969;
CC Evidence={ECO:0000305|PubMed:3061453};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000269|PubMed:15683236, ECO:0000269|PubMed:22342614,
CC ECO:0000269|PubMed:3061453, ECO:0000269|PubMed:8508789};
CC Note=Binds 1 FMN per subunit. {ECO:0000269|PubMed:15683236,
CC ECO:0000269|PubMed:22342614, ECO:0000269|PubMed:3061453};
CC -!- ACTIVITY REGULATION: Is inhibited in vitro by CCPST (4-carboxy-5-(4-
CC chlorophenyl)sulfanyl-1,2,3-thiadiazole).
CC {ECO:0000269|PubMed:22342614}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.046 mM for L-2-hydroxyoctanoate (with DCIP as electron acceptor)
CC {ECO:0000269|PubMed:15683236};
CC KM=1.36 mM for L-2-hydroxyhexadecanoate (with DCIP as electron
CC acceptor) {ECO:0000269|PubMed:15683236};
CC KM=0.69 mM for DL-2-hydroxybutanoate (with DCIP as electron acceptor)
CC {ECO:0000269|PubMed:8508789};
CC KM=0.6 mM for L-2-hydroxybutanoate (with DCIP as electron acceptor)
CC {ECO:0000269|PubMed:3061453};
CC KM=0.25 mM for L-2-hydroxyhexanoate (with DCIP as electron acceptor)
CC {ECO:0000269|PubMed:3061453};
CC KM=0.8 mM for L-mandelate (with DCIP as electron acceptor)
CC {ECO:0000269|PubMed:3061453};
CC Vmax=12 umol/min/mg enzyme with 2-hydroxybutanoate as substrate and
CC DCIP as electron acceptor {ECO:0000269|PubMed:3061453};
CC Vmax=5.8 umol/min/mg enzyme with 2-hydroxyhexanoate as substrate and
CC DCIP as electron acceptor {ECO:0000269|PubMed:3061453};
CC Vmax=21 umol/min/mg enzyme with L-mandelate as substrate and DCIP as
CC electron acceptor {ECO:0000269|PubMed:3061453};
CC Note=kcat is 0.99 sec(-1) for the oxidation of L-2-hydroxyoctanoate
CC with DCIP instead of O2 as electron acceptor. kcat is 0.34 sec(-1)
CC for the oxidation of L-2-hydroxyhexadecanoate with DCIP instead of O2
CC as electron acceptor (PubMed:15683236). kcat is 1.66 sec(-1) for the
CC oxidation of DL-2-hydroxybutanoate with DCIP instead of O2 as
CC electron acceptor (PubMed:8508789). {ECO:0000269|PubMed:15683236,
CC ECO:0000269|PubMed:8508789};
CC -!- SUBUNIT: Homotetramer (PubMed:8508789). Could also form homooctamer.
CC {ECO:0000269|PubMed:15683236, ECO:0000269|PubMed:8508789}.
CC -!- SUBCELLULAR LOCATION: Peroxisome.
CC -!- TISSUE SPECIFICITY: Expressed in kidney. {ECO:0000269|PubMed:1939137,
CC ECO:0000269|PubMed:8508789}.
CC -!- SIMILARITY: Belongs to the FMN-dependent alpha-hydroxy acid
CC dehydrogenase family. {ECO:0000255|PROSITE-ProRule:PRU00683}.
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DR EMBL; X67156; CAA47629.1; -; mRNA.
DR EMBL; BC078781; AAH78781.1; -; mRNA.
DR PIR; S33322; S33322.
DR RefSeq; NP_114471.1; NM_032082.2.
DR PDB; 1TB3; X-ray; 2.30 A; A/B/C/D/E/F/G/H=2-353.
DR PDB; 3SGZ; X-ray; 1.35 A; A/B/C=2-353.
DR PDBsum; 1TB3; -.
DR PDBsum; 3SGZ; -.
DR AlphaFoldDB; Q07523; -.
DR SMR; Q07523; -.
DR IntAct; Q07523; 1.
DR STRING; 10116.ENSRNOP00000040223; -.
DR BindingDB; Q07523; -.
DR ChEMBL; CHEMBL2021745; -.
DR iPTMnet; Q07523; -.
DR PhosphoSitePlus; Q07523; -.
DR PaxDb; Q07523; -.
DR PRIDE; Q07523; -.
DR Ensembl; ENSRNOT00000046942; ENSRNOP00000040223; ENSRNOG00000019470.
DR GeneID; 84029; -.
DR KEGG; rno:84029; -.
DR UCSC; RGD:70972; rat.
DR CTD; 51179; -.
DR RGD; 70972; Hao2.
DR eggNOG; KOG0538; Eukaryota.
DR GeneTree; ENSGT00390000018717; -.
DR HOGENOM; CLU_020639_6_1_1; -.
DR InParanoid; Q07523; -.
DR OrthoDB; 879633at2759; -.
DR PhylomeDB; Q07523; -.
DR TreeFam; TF313363; -.
DR BRENDA; 1.1.3.15; 5301.
DR Reactome; R-RNO-390918; Peroxisomal lipid metabolism.
DR Reactome; R-RNO-9033241; Peroxisomal protein import.
DR SABIO-RK; Q07523; -.
DR EvolutionaryTrace; Q07523; -.
DR PRO; PR:Q07523; -.
DR Proteomes; UP000002494; Chromosome 2.
DR Bgee; ENSRNOG00000019470; Expressed in adult mammalian kidney and 15 other tissues.
DR ExpressionAtlas; Q07523; baseline and differential.
DR Genevisible; Q07523; RN.
DR GO; GO:0005782; C:peroxisomal matrix; ISO:RGD.
DR GO; GO:0005777; C:peroxisome; IDA:HGNC-UCL.
DR GO; GO:0003973; F:(S)-2-hydroxy-acid oxidase activity; IDA:RGD.
DR GO; GO:0010181; F:FMN binding; IDA:RGD.
DR GO; GO:0042802; F:identical protein binding; IPI:RGD.
DR GO; GO:0019395; P:fatty acid oxidation; ISO:RGD.
DR GO; GO:0018924; P:mandelate metabolic process; IDA:RGD.
DR CDD; cd02809; alpha_hydroxyacid_oxid_FMN; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR012133; Alpha-hydoxy_acid_DH_FMN.
DR InterPro; IPR000262; FMN-dep_DH.
DR InterPro; IPR037396; FMN_HAD.
DR InterPro; IPR008259; FMN_hydac_DH_AS.
DR Pfam; PF01070; FMN_dh; 1.
DR PIRSF; PIRSF000138; Al-hdrx_acd_dh; 1.
DR PROSITE; PS00557; FMN_HYDROXY_ACID_DH_1; 1.
DR PROSITE; PS51349; FMN_HYDROXY_ACID_DH_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Fatty acid metabolism;
KW Flavoprotein; FMN; Lipid metabolism; Oxidoreductase; Peroxisome;
KW Phosphoprotein; Reference proteome.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:1939137,
FT ECO:0000269|PubMed:3061453"
FT CHAIN 2..353
FT /note="2-Hydroxyacid oxidase 2"
FT /id="PRO_0000206322"
FT DOMAIN 2..353
FT /note="FMN hydroxy acid dehydrogenase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT MOTIF 351..353
FT /note="Microbody targeting signal"
FT /evidence="ECO:0000255"
FT ACT_SITE 248
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683,
FT ECO:0000305|PubMed:15683236"
FT BINDING 77..79
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT BINDING 106
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT BINDING 128
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT BINDING 130
FT /ligand="a 2-oxocarboxylate"
FT /ligand_id="ChEBI:CHEBI:35179"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT BINDING 156
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT BINDING 165
FT /ligand="a 2-oxocarboxylate"
FT /ligand_id="ChEBI:CHEBI:35179"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT BINDING 224
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT BINDING 251
FT /ligand="a 2-oxocarboxylate"
FT /ligand_id="ChEBI:CHEBI:35179"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT BINDING 279..283
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT BINDING 302..303
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:15683236,
FT ECO:0000269|PubMed:22342614, ECO:0007744|PDB:1TB3,
FT ECO:0007744|PDB:3SGZ"
FT MOD_RES 133
FT /note="Phosphoserine"
FT /evidence="ECO:0007744|PubMed:22673903"
FT HELIX 6..15
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 19..26
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 33..43
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 62..64
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 67..75
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 81..83
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 88..99
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 102..105
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 113..119
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 124..128
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 135..147
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 153..156
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 166..174
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 177..180
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 207..216
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 221..226
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 229..237
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 241..245
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 248..250
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 259..270
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 273..281
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 285..293
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 297..302
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 303..335
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 340..342
FT /evidence="ECO:0007829|PDB:3SGZ"
FT HELIX 345..347
FT /evidence="ECO:0007829|PDB:3SGZ"
FT STRAND 348..350
FT /evidence="ECO:0007829|PDB:3SGZ"
SQ SEQUENCE 353 AA; 39201 MW; A2FDD4BF34E269E5 CRC64;
MPLVCLADFK AHAQKQLSKT SWDFIEGEAD DGITYSENIA AFKRIRLRPR YLRDMSKVDT
RTTIQGQEIS APICISPTAF HSIAWPDGEK STARAAQEAN ICYVISSYAS YSLEDIVAAA
PEGFRWFQLY MKSDWDFNKQ MVQRAEALGF KALVITIDTP VLGNRRRDKR NQLNLEANIL
LKDLRALKEE KPTQSVPVSF PKASFCWNDL SLLQSITRLP IILKGILTKE DAELAMKHNV
QGIVVSNHGG RQLDEVSASI DALREVVAAV KGKIEVYMDG GVRTGTDVLK ALALGARCIF
LGRPILWGLA CKGEDGVKEV LDILTAELHR CMTLSGCQSV AEISPDLIQF SRL
