OFOA_SULSP
ID OFOA_SULSP Reviewed; 632 AA.
AC P72578;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 03-AUG-2022, entry version 76.
DE RecName: Full=2-oxoacid:ferredoxin oxidoreductase subunit alpha {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;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-23, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBCELLULAR LOCATION,
RP SUBUNIT, SUBSTRATE SPECIFICITY, AND REACTION MECHANISM.
RC STRAIN=7 {ECO:0000312|EMBL:BAA10898.1};
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, MUTAGENESIS OF
RP TYR-253; PRO-254; ILE-255; THR-256 AND PRO-257, SUBCELLULAR LOCATION,
RP 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 THR-256; ARG-344 AND THR-353, 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 -!- 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 -!- DOMAIN: The Tyr-Pro-Ile-Thr-Pro (YPITP) motif is important for the
CC turnover of the reaction, presumably through its flexibility and
CC mobility. {ECO:0000269|PubMed:8902625}.
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; D64024; BAA10898.1; -; Genomic_DNA.
DR PIR; JC4919; JC4919.
DR AlphaFoldDB; P72578; -.
DR SMR; P72578; -.
DR KEGG; ag:BAA10898; -.
DR BioCyc; MetaCyc:MON-11911; -.
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:0019164; F:pyruvate synthase activity; IDA:UniProtKB.
DR CDD; cd07034; TPP_PYR_PFOR_IOR-alpha_like; 1.
DR Gene3D; 3.40.50.920; -; 1.
DR Gene3D; 3.40.920.10; -; 1.
DR InterPro; IPR022367; 2-oxoacid/accept_OxRdtase_asu.
DR InterPro; IPR019752; Pyrv/ketoisovalerate_OxRed_cat.
DR InterPro; IPR002880; Pyrv_Fd/Flavodoxin_OxRdtase_N.
DR InterPro; IPR002869; Pyrv_flavodox_OxRed_cen.
DR InterPro; IPR029061; THDP-binding.
DR InterPro; IPR009014; Transketo_C/PFOR_II.
DR Pfam; PF01558; POR; 1.
DR Pfam; PF01855; POR_N; 1.
DR SUPFAM; SSF52518; SSF52518; 1.
DR SUPFAM; SSF52922; SSF52922; 1.
DR SUPFAM; SSF53323; SSF53323; 1.
DR TIGRFAMs; TIGR03710; OAFO_sf; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Direct protein sequencing; Oxidoreductase; Pyruvate.
FT CHAIN 1..632
FT /note="2-oxoacid:ferredoxin oxidoreductase subunit alpha"
FT /id="PRO_0000445529"
FT MOTIF 253..257
FT /note="YPITP motif"
FT /evidence="ECO:0000269|PubMed:11683888"
FT BINDING 256
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q96XT2"
FT BINDING 344
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q96XT2"
FT SITE 353
FT /note="Plays a key role in the broad substrate specificity"
FT /evidence="ECO:0000269|PubMed:12009405"
FT MUTAGEN 253
FT /note="Y->A,W: Loss of oxidoreductase activity."
FT /evidence="ECO:0000269|PubMed:11683888"
FT MUTAGEN 253
FT /note="Y->F: Significant decrease in the catalytic
FT efficiency, while the affinity for 2-oxoacid is only
FT slightly affected."
FT /evidence="ECO:0000269|PubMed:11683888"
FT MUTAGEN 254
FT /note="P->G: Significant decrease in the catalytic
FT efficiency, while the affinity for 2-oxoacid is only
FT slightly affected."
FT /evidence="ECO:0000269|PubMed:11683888"
FT MUTAGEN 255
FT /note="I->L,M,S,V: Significant decrease in the catalytic
FT efficiency, while the affinity for 2-oxoacid is only
FT slightly affected."
FT /evidence="ECO:0000269|PubMed:11683888"
FT MUTAGEN 256
FT /note="T->A,S,V: Significant decrease in the catalytic
FT efficiency, while the affinity for 2-oxoacid is only
FT slightly affected."
FT /evidence="ECO:0000269|PubMed:11683888,
FT ECO:0000269|PubMed:12009405"
FT MUTAGEN 257
FT /note="P->A,G,V: Loss of oxidoreductase activity."
FT /evidence="ECO:0000269|PubMed:11683888"
FT MUTAGEN 344
FT /note="R->A,K: Loss of oxidoreductase activity."
FT /evidence="ECO:0000269|PubMed:12009405"
FT MUTAGEN 353
FT /note="T->I,V: Strong decrease of the oxidoreductase
FT activity with pyruvate, 2-oxobutyrate and 2-oxoglutarate."
FT /evidence="ECO:0000269|PubMed:12009405"
SQ SEQUENCE 632 AA; 70759 MW; 362566F1D59D72D9 CRC64;
MRLSWVIGGA QGTGIDTAAN IFGNAVASAG YYIYGNREYY SNIKGGHSYF SLTISDKRVR
SNTQKIDILV SFDAETVFQH FYDVKDILIY NKAVETTKID AVQSMEPELA ERIKDFLTKQ
GYETTVKGAL EYASKNNVTL IPVNYDEIAK KVADEMKVPL SVTERVKNIV GITISYKLLG
LDVNYLIEAI NSTFKQDLYR KMNELAVKDS YDIVESRYNL KPSSKERRRF WLDGNTAVAI
GKIYGGVRFQ SYYPITPASD ESVYIEAHQD VLMEDPITGD KKKGTIVVVQ AEDELAAINM
AIGAALTGVR AATATSGPGF SLMVEGLGWA GMNEVPVVIT YYIRGGPSTG LPTRTAQSDL
IFPIFAGHGE FPKIVLASGD HAEAFKDAIW ALNLAEKYQT PVIHLVEKTL ANSYSTIPYE
ELELDKLKAE RGKIVESGDI SYKRFKFTED GISPRAFLGK ATMYYTGDEH NEEGHISEDV
VNRTMMYEKR MKKLEVADKE IPEESRVKIY GDLNSRNLII TWGSPTGVLR DILEESNFDF
TLLQIRMFSP FPKNLVSKLM EGRDKIITVE GNYLAQTSLL VKMYTGKDVT NSILKWNGRP
FLRDELEEAL IKVIKDGEKR VVLNGGIYTS ME
