GOX_SPIOL
ID GOX_SPIOL Reviewed; 369 AA.
AC P05414;
DT 01-NOV-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1988, sequence version 1.
DT 03-AUG-2022, entry version 138.
DE RecName: Full=Glycolate oxidase {ECO:0000303|PubMed:1324737, ECO:0000303|PubMed:2824469, ECO:0000303|PubMed:3286256};
DE Short=GAO {ECO:0000303|PubMed:1324737};
DE Short=GOX {ECO:0000303|PubMed:9144771};
DE EC=1.1.3.15 {ECO:0000269|PubMed:1324737, ECO:0000269|PubMed:7705356};
DE AltName: Full=Short chain alpha-hydroxy acid oxidase;
OS Spinacia oleracea (Spinach).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC Caryophyllales; Chenopodiaceae; Chenopodioideae; Anserineae; Spinacia.
OX NCBI_TaxID=3562;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=2824469; DOI=10.1016/s0021-9258(18)47662-4;
RA Volokita M., Somerville C.R.;
RT "The primary structure of spinach glycolate oxidase deduced from the DNA
RT sequence of a cDNA clone.";
RL J. Biol. Chem. 262:15825-15828(1987).
RN [2]
RP PROTEIN SEQUENCE, AND ACETYLATION AT MET-1.
RX PubMed=3286256; DOI=10.1111/j.1432-1033.1988.tb14029.x;
RA Cederlund E., Lindqvist Y., Soederlund G., Braenden C.-I., Joernvall H.;
RT "Primary structure of glycolate oxidase from spinach.";
RL Eur. J. Biochem. 173:523-530(1988).
RN [3]
RP FUNCTION, COFACTOR, AND ACTIVE SITE.
RX PubMed=2644287; DOI=10.1016/s0021-9258(18)94112-8;
RA Lindqvist Y., Braenden C.-I.;
RT "The active site of spinach glycolate oxidase.";
RL J. Biol. Chem. 264:3624-3628(1989).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=1324737; DOI=10.1016/0167-4781(92)90046-3;
RA Macheroux P., Mulrooney S.B., Williams C.H. Jr., Massey V.;
RT "Direct expression of active spinach glycolate oxidase in Escherichia
RT coli.";
RL Biochim. Biophys. Acta 1132:11-16(1992).
RN [5] {ECO:0007744|PDB:1GOX}
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEX WITH FMN, AND COFACTOR.
RX PubMed=2681790; DOI=10.1016/0022-2836(89)90178-2;
RA Lindqvist Y.;
RT "Refined structure of spinach glycolate oxidase at 2-A resolution.";
RL J. Mol. Biol. 209:151-166(1989).
RN [6] {ECO:0007744|PDB:1GYL}
RP X-RAY CRYSTALLOGRAPHY (3.00 ANGSTROMS) OF MUTANT PHE-24 IN COMPLEX WITH
RP FMN, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE
RP SPECIFICITY, COFACTOR, AND MUTAGENESIS OF TYR-24 AND TRP-108.
RX PubMed=7705356; DOI=10.1111/j.1432-1033.1995.tb20278.x;
RA Stenberg K., Clausen T., Lindqvist Y., Macheroux P.;
RT "Involvement of Tyr24 and Trp108 in substrate binding and substrate
RT specificity of glycolate oxidase.";
RL Eur. J. Biochem. 228:408-416(1995).
RN [7] {ECO:0007744|PDB:1AL7, ECO:0007744|PDB:1AL8}
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 1-359 IN COMPLEXES WITH FMN AND
RP INHIBITORS, COFACTOR, AND SUBUNIT.
RX PubMed=9144771; DOI=10.1002/pro.5560060506;
RA Stenberg K., Lindqvist Y.;
RT "Three-dimensional structures of glycolate oxidase with bound active-site
RT inhibitors.";
RL Protein Sci. 6:1009-1015(1997).
CC -!- FUNCTION: Catalyzes the oxidation of glycolate to glyoxylate, with a
CC reduction of O2 to H2O2 (PubMed:1324737, PubMed:7705356). Is a key
CC enzyme in photorespiration in green plants (Probable). To a lesser
CC extent, is also able to use L-lactate and 2-hydroxbyutanoate as
CC substrate in vitro, but shows almost no activity with L-mandelate
CC (PubMed:7705356). {ECO:0000269|PubMed:1324737,
CC ECO:0000269|PubMed:7705356, ECO:0000305|PubMed:2644287}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=glycolate + O2 = glyoxylate + H2O2; Xref=Rhea:RHEA:25311,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16240, ChEBI:CHEBI:29805,
CC ChEBI:CHEBI:36655; EC=1.1.3.15; Evidence={ECO:0000269|PubMed:1324737,
CC ECO:0000269|PubMed:7705356};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:25312;
CC Evidence={ECO:0000305|PubMed:7705356};
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:0000269|PubMed:7705356};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:16790;
CC Evidence={ECO:0000305|PubMed:7705356};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000269|PubMed:2644287, ECO:0000269|PubMed:2681790,
CC ECO:0000269|PubMed:7705356, ECO:0000269|PubMed:9144771};
CC Note=Binds 1 FMN per subunit. {ECO:0000269|PubMed:2681790};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.2 mM for glycolate (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:1324737};
CC KM=1 mM for glycolate {ECO:0000269|PubMed:7705356};
CC KM=11.5 mM for L-lactate {ECO:0000269|PubMed:7705356};
CC KM=0.21 mM for O2 {ECO:0000269|PubMed:7705356};
CC KM=3 mM for L-2-hydroxbyutanoate {ECO:0000269|PubMed:7705356};
CC KM=58 mM for L-mandelate {ECO:0000269|PubMed:7705356};
CC Note=kcat is 20 sec(-1) with glycolate as substrate. kcat is 11 sec(-
CC 1) with L-lactate as substrate. kcat is 13.2 sec(-1) with L-2-
CC hydroxbyutanoate as substrate. kcat is 0.09 sec(-1) with L-mandelate
CC as substrate. {ECO:0000269|PubMed:7705356};
CC -!- PATHWAY: Photosynthesis; photorespiration; glycine from 2-
CC phosphoglycolate: step 2/3. {ECO:0000305|PubMed:2644287}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000305|PubMed:9144771}.
CC -!- SUBCELLULAR LOCATION: Peroxisome.
CC -!- SIMILARITY: Belongs to the FMN-dependent alpha-hydroxy acid
CC dehydrogenase family. {ECO:0000255|PROSITE-ProRule:PRU00683}.
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DR EMBL; J03492; AAA34030.1; -; mRNA.
DR PIR; A28496; OXSPH.
DR PDB; 1AL7; X-ray; 2.60 A; A=1-359.
DR PDB; 1AL8; X-ray; 2.20 A; A=1-359.
DR PDB; 1GOX; X-ray; 2.00 A; A=1-369.
DR PDB; 1GYL; X-ray; 3.00 A; A/B=1-369.
DR PDB; 1HUV; X-ray; 2.15 A; A=176-195.
DR PDB; 1P4C; X-ray; 1.35 A; A=176-195.
DR PDB; 1P5B; X-ray; 1.35 A; A=176-195.
DR PDB; 2A7N; X-ray; 1.80 A; A=176-195.
DR PDB; 2A7P; X-ray; 2.20 A; A=176-195.
DR PDB; 2A85; X-ray; 2.50 A; A=176-195.
DR PDBsum; 1AL7; -.
DR PDBsum; 1AL8; -.
DR PDBsum; 1GOX; -.
DR PDBsum; 1GYL; -.
DR PDBsum; 1HUV; -.
DR PDBsum; 1P4C; -.
DR PDBsum; 1P5B; -.
DR PDBsum; 2A7N; -.
DR PDBsum; 2A7P; -.
DR PDBsum; 2A85; -.
DR AlphaFoldDB; P05414; -.
DR SMR; P05414; -.
DR iPTMnet; P05414; -.
DR PRIDE; P05414; -.
DR OrthoDB; 879633at2759; -.
DR UniPathway; UPA00951; UER00912.
DR EvolutionaryTrace; P05414; -.
DR GO; GO:0005777; C:peroxisome; IEA:UniProtKB-SubCell.
DR GO; GO:0003973; F:(S)-2-hydroxy-acid oxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR GO; GO:0009854; P:oxidative photosynthetic carbon pathway; IEA:UniProtKB-UniPathway.
DR GO; GO:0051707; P:response to other organism; IEA:UniProt.
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; Acetylation; Direct protein sequencing; Flavoprotein; FMN;
KW Glycolate pathway; Oxidoreductase; Peroxisome; Photorespiration.
FT CHAIN 1..369
FT /note="Glycolate oxidase"
FT /id="PRO_0000206325"
FT DOMAIN 1..359
FT /note="FMN hydroxy acid dehydrogenase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT MOTIF 367..369
FT /note="Microbody targeting signal"
FT /evidence="ECO:0000255"
FT ACT_SITE 254
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00683,
FT ECO:0000305|PubMed:2644287"
FT BINDING 24
FT /ligand="glyoxylate"
FT /ligand_id="ChEBI:CHEBI:36655"
FT /evidence="ECO:0000250|UniProtKB:Q9UJM8"
FT BINDING 77..79
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT BINDING 106
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:9144771,
FT ECO:0007744|PDB:1AL7"
FT BINDING 127..129
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT BINDING 129
FT /ligand="glyoxylate"
FT /ligand_id="ChEBI:CHEBI:36655"
FT /evidence="ECO:0000250|UniProtKB:Q9UJM8"
FT BINDING 155
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT BINDING 164
FT /ligand="glyoxylate"
FT /ligand_id="ChEBI:CHEBI:36655"
FT /evidence="ECO:0000250|UniProtKB:Q9UJM8"
FT BINDING 230
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT BINDING 252
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT BINDING 254
FT /ligand="glyoxylate"
FT /ligand_id="ChEBI:CHEBI:36655"
FT /evidence="ECO:0000250|UniProtKB:Q9UJM8"
FT BINDING 257
FT /ligand="glyoxylate"
FT /ligand_id="ChEBI:CHEBI:36655"
FT /evidence="ECO:0000250|UniProtKB:Q9UJM8"
FT BINDING 285..289
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT BINDING 308..309
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:2681790,
FT ECO:0000269|PubMed:9144771, ECO:0007744|PDB:1AL7,
FT ECO:0007744|PDB:1GOX"
FT SITE 108
FT /note="Involved in determining the substrate specificity of
FT glycolate oxidase"
FT /evidence="ECO:0000305|PubMed:7705356"
FT MOD_RES 1
FT /note="N-acetylmethionine"
FT /evidence="ECO:0000269|PubMed:3286256"
FT MUTAGEN 24
FT /note="Y->F: 10-fold decrease in affinity for glycolate."
FT /evidence="ECO:0000269|PubMed:7705356"
FT MUTAGEN 108
FT /note="W->S: 100-fold decrease in affinity for glycolate
FT and 500-fold decrease in activity."
FT /evidence="ECO:0000269|PubMed:7705356"
FT HELIX 8..16
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 19..26
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 33..40
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 41..44
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 45..47
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 62..64
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 67..75
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 81..83
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 88..98
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 103..105
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 113..117
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 124..128
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 131..133
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 134..146
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 151..155
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 165..169
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:1P4C"
FT STRAND 184..186
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 199..205
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 213..222
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 227..230
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 235..243
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 247..251
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 254..256
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 260..262
FT /evidence="ECO:0007829|PDB:1GYL"
FT HELIX 265..275
FT /evidence="ECO:0007829|PDB:1GOX"
FT TURN 276..278
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 282..287
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 291..300
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 303..307
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 309..341
FT /evidence="ECO:0007829|PDB:1GOX"
FT TURN 346..348
FT /evidence="ECO:0007829|PDB:1GOX"
FT HELIX 351..353
FT /evidence="ECO:0007829|PDB:1GOX"
FT STRAND 354..356
FT /evidence="ECO:0007829|PDB:1GOX"
SQ SEQUENCE 369 AA; 40286 MW; 892F1B3D0C1B48E0 CRC64;
MEITNVNEYE AIAKQKLPKM VYDYYASGAE DQWTLAENRN AFSRILFRPR ILIDVTNIDM
TTTILGFKIS MPIMIAPTAM QKMAHPEGEY ATARAASAAG TIMTLSSWAT SSVEEVASTG
PGIRFFQLYV YKDRNVVAQL VRRAERAGFK AIALTVDTPR LGRREADIKN RFVLPPFLTL
KNFEGIDLGK MDKANDSGLS SYVAGQIDRS LSWKDVAWLQ TITSLPILVK GVITAEDARL
AVQHGAAGII VSNHGARQLD YVPATIMALE EVVKAAQGRI PVFLDGGVRR GTDVFKALAL
GAAGVFIGRP VVFSLAAEGE AGVKKVLQMM RDEFELTMAL SGCRSLKEIS RSHIAADWDG
PSSRAVARL
