OFOB_SULSP
ID OFOB_SULSP Reviewed; 305 AA.
AC P72579;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 03-AUG-2022, entry version 59.
DE RecName: Full=2-oxoacid:ferredoxin oxidoreductase subunit beta {ECO:0000303|PubMed:8902625};
DE Short=OFOR {ECO:0000303|PubMed:11683888};
DE EC=1.2.7.11 {ECO:0000269|PubMed:11683888, ECO:0000269|PubMed:12009405, ECO:0000269|PubMed:8902625};
OS Sulfolobus sp.
OC Archaea; Crenarchaeota; Thermoprotei; Sulfolobales; Sulfolobaceae;
OC Sulfolobus; unclassified Sulfolobus.
OX NCBI_TaxID=2288 {ECO:0000312|EMBL:BAA10899.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, MASS SPECTROMETRY, SUBCELLULAR
RP LOCATION, SUBUNIT, SUBSTRATE SPECIFICITY, AND REACTION MECHANISM.
RC STRAIN=7 {ECO:0000312|EMBL:BAA10899.1};
RX PubMed=8902625; DOI=10.1093/oxfordjournals.jbchem.a021454;
RA Zhang Q., Iwasaki T., Wakagi T., Oshima T.;
RT "2-oxoacid:ferredoxin oxidoreductase from the thermoacidophilic archaeon,
RT Sulfolobus sp. strain 7.";
RL J. Biochem. 120:587-599(1996).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP SUBCELLULAR LOCATION, SUBUNIT, AND SUBSTRATE SPECIFICITY.
RC STRAIN=7;
RX PubMed=11683888; DOI=10.1046/j.1432-1033.2001.02504.x;
RA Fukuda E., Kino H., Matsuzawa H., Wakagi T.;
RT "Role of a highly conserved YPITP motif in 2-oxoacid:ferredoxin
RT oxidoreductase: heterologous expression of the gene from Sulfolobus
RT sp.strain 7, and characterization of the recombinant and variant enzymes.";
RL Eur. J. Biochem. 268:5639-5646(2001).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP LYS-49 AND LEU-123, SUBUNIT, AND SUBSTRATE SPECIFICITY.
RX PubMed=12009405; DOI=10.1016/s0167-4838(02)00280-7;
RA Fukuda E., Wakagi T.;
RT "Substrate recognition by 2-oxoacid:ferredoxin oxidoreductase from
RT Sulfolobus sp. strain 7.";
RL Biochim. Biophys. Acta 1597:74-80(2002).
CC -!- FUNCTION: Catalyzes the coenzyme A-dependent oxidative decarboxylation
CC of different 2-oxoacids such as 2-oxoglutarate, pyruvate and 2-
CC oxobutyrate to form their CoA derivatives.
CC {ECO:0000269|PubMed:11683888, ECO:0000269|PubMed:12009405,
CC ECO:0000269|PubMed:8902625}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 2-oxocarboxylate + CoA + 2 oxidized [2Fe-2S]-[ferredoxin] =
CC an acyl-CoA + CO2 + H(+) + 2 reduced [2Fe-2S]-[ferredoxin];
CC Xref=Rhea:RHEA:42316, Rhea:RHEA-COMP:10000, Rhea:RHEA-COMP:10001,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:16526, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738, ChEBI:CHEBI:35179, ChEBI:CHEBI:57287,
CC ChEBI:CHEBI:58342; EC=1.2.7.11;
CC Evidence={ECO:0000269|PubMed:11683888, ECO:0000269|PubMed:12009405,
CC ECO:0000269|PubMed:8902625};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000269|PubMed:8902625, ECO:0000305|PubMed:11683888};
CC Note=Binds 1 [4Fe-4S] cluster per subunit.
CC {ECO:0000269|PubMed:8902625};
CC -!- COFACTOR:
CC Name=thiamine diphosphate; Xref=ChEBI:CHEBI:58937;
CC Evidence={ECO:0000269|PubMed:8902625, ECO:0000305|PubMed:11683888};
CC Note=Binds 1 thiamine pyrophosphate per subunit.
CC {ECO:0000269|PubMed:8902625};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:8902625, ECO:0000305|PubMed:11683888};
CC Note=Binds 1 Mg(2+) per subunit. {ECO:0000269|PubMed:8902625};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=250 uM for pyruvate (at pH 6.8 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:8902625};
CC KM=280 uM for pyruvate {ECO:0000269|PubMed:12009405};
CC KM=480 uM for 2-oxobutyrate {ECO:0000269|PubMed:12009405};
CC KM=870 uM for 2-oxoglutarate (at pH 6.8 and 50 degrees Celsius)
CC {ECO:0000269|PubMed:12009405, ECO:0000269|PubMed:8902625};
CC Vmax=86 umol/min/mg enzyme with 2-oxoglutarate as substrate (at pH
CC 6.8 and 50 degrees Celsius) {ECO:0000269|PubMed:8902625};
CC Vmax=30 umol/min/mg enzyme with pyruvate as substrate
CC {ECO:0000269|PubMed:12009405};
CC Vmax=28 umol/min/mg enzyme with 2-oxobutyrate as substrate
CC {ECO:0000269|PubMed:12009405};
CC Vmax=11 umol/min/mg enzyme with 2-oxoglutarate as substrate
CC {ECO:0000269|PubMed:12009405};
CC Note=kcat is 51 sec(-1) for pyruvate as substrate (PubMed:11683888).
CC kcat is 19 sec(-1) for 2-oxoglutarate as substrate (PubMed:11683888).
CC {ECO:0000269|PubMed:11683888};
CC pH dependence:
CC Optimum pH is 8.5. {ECO:0000269|PubMed:11683888};
CC Temperature dependence:
CC Optimum temperature is 90 degrees Celsius.
CC {ECO:0000269|PubMed:11683888};
CC -!- SUBUNIT: Heterodimer composed of an alpha and a beta subunit.
CC {ECO:0000269|PubMed:11683888, ECO:0000269|PubMed:12009405,
CC ECO:0000269|PubMed:8902625}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:11683888,
CC ECO:0000269|PubMed:8902625}.
CC -!- MASS SPECTROMETRY: Mass=33566; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:8902625};
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DR EMBL; D64024; BAA10899.1; -; Genomic_DNA.
DR PIR; JC4920; JC4920.
DR AlphaFoldDB; P72579; -.
DR SMR; P72579; -.
DR KEGG; ag:BAA10899; -.
DR BioCyc; MetaCyc:MON-11912; -.
DR BRENDA; 1.2.7.11; 6164.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0018491; F:2-oxobutyrate synthase activity; IDA:UniProtKB.
DR GO; GO:0047553; F:2-oxoglutarate synthase activity; IDA:UniProtKB.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; ISS:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; ISS:UniProtKB.
DR GO; GO:0019164; F:pyruvate synthase activity; IDA:UniProtKB.
DR GO; GO:0030976; F:thiamine pyrophosphate binding; ISS:UniProtKB.
DR InterPro; IPR011896; OFOB.
DR InterPro; IPR032686; PFO_beta_C.
DR InterPro; IPR029061; THDP-binding.
DR InterPro; IPR011766; TPP_enzyme-bd_C.
DR Pfam; PF12367; PFO_beta_C; 1.
DR Pfam; PF02775; TPP_enzyme_C; 1.
DR SUPFAM; SSF52518; SSF52518; 1.
DR TIGRFAMs; TIGR02177; PorB_KorB; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Cytoplasm; Iron; Iron-sulfur; Magnesium; Metal-binding;
KW Oxidoreductase; Thiamine pyrophosphate.
FT CHAIN 1..305
FT /note="2-oxoacid:ferredoxin oxidoreductase subunit beta"
FT /id="PRO_0000445530"
FT BINDING 12
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 15
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 44..47
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 46
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 65
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 90
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 91..92
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 118
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 120
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 122..123
FT /ligand="thiamine diphosphate"
FT /ligand_id="ChEBI:CHEBI:58937"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT BINDING 197
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q96XT4"
FT SITE 125
FT /note="Plays an important role in the binding of CoA"
FT /evidence="ECO:0000250|UniProtKB:Q96Y68"
FT MUTAGEN 49
FT /note="K->I: Strong decrease of the oxidoreductase activity
FT with pyruvate, 2-oxobutyrate and 2-oxoglutarate."
FT /evidence="ECO:0000269|PubMed:12009405"
FT MUTAGEN 49
FT /note="K->R: Increase the oxidoreductase activity with
FT pyruvate."
FT /evidence="ECO:0000269|PubMed:12009405"
FT MUTAGEN 49
FT /note="K->V: Slight decrease of the oxidoreductase activity
FT with pyruvate, 2-oxobutyrate and 2-oxoglutarate."
FT /evidence="ECO:0000269|PubMed:12009405"
FT MUTAGEN 123
FT /note="L->A,I: Strong decrease of the oxidoreductase
FT activity with pyruvate, 2-oxobutyrate and 2-oxoglutarate."
FT /evidence="ECO:0000269|PubMed:12009405"
FT MUTAGEN 123
FT /note="L->N: Strong decrease of the oxidoreductase activity
FT with pyruvate and 2-oxobutyrate. However, this mutant shows
FT almost the same activity with 2-oxoglutarate as the wild-
FT type."
FT /evidence="ECO:0000269|PubMed:12009405"
SQ SEQUENCE 305 AA; 33609 MW; 4DF4E3596BDCC578 CRC64;
MAAFTPQWND WCPGCGNFGI LNAEQQAIVE LGVDTKNVVV VSGIGCSGKI PHFFRTPISG
VHTLHGRAIA FATGIKLSNP DLVVIVNGGD GDLLGIGAGH FVAAGRRNVD MVVILHDNGV
YGLTKGQASP TLKRGEKPKS LPRPNINDAV NPIALAISSG YTFVARGYAY DVKHLKELIK
SAIKHKGLAL IDVLQPCPTY NDINTKEWYD KRIYKLDTLP DWDPVVKKPE EVNEKIKRAI
DKSLEWGDRI PIGIFYQNEL VPSYEERIKA NSPAYLDYTP AKQLIEKEGK LTTIIDPLLK
EREVD
