ID PNO_EUGGR Reviewed; 1803 AA.
AC Q94IN5; Q9FZP8;
DT 19-JUL-2003, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 27-MAR-2024, entry version 125.
DE RecName: Full=Pyruvate dehydrogenase [NADP(+)], mitochondrial;
DE EC=1.2.1.51 {ECO:0000269|PubMed:3110154};
DE AltName: Full=Aquacobalamin reductase [NADPH];
DE AltName: Full=EgPNOmt;
DE AltName: Full=Pyruvate:NADP(+) oxidoreductase;
DE Flags: Precursor;
GN Name=PNO;
OS Euglena gracilis.
OC Eukaryota; Discoba; Euglenozoa; Euglenida; Spirocuta; Euglenophyceae;
OC Euglenales; Euglenaceae; Euglena.
OX NCBI_TaxID=3039 {ECO:0000312|EMBL:CAC37628.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RX PubMed=11023353; DOI=10.1016/s0014-5793(00)01882-2;
RA Nakazawa M., Inui H., Yamaji R., Yamamoto T., Takenaka S., Ueda M.,
RA Nakano Y., Miyatake K.;
RT "The origin of pyruvate:NADP+ oxidoreductase in mitochondria of Euglena
RT gracilis.";
RL FEBS Lett. 479:155-156(2000).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], SUBCELLULAR LOCATION, AND TISSUE SPECIFICITY.
RC STRAIN=SAG1224-5/25;
RX PubMed=11319255; DOI=10.1093/oxfordjournals.molbev.a003853;
RA Rotte C., Stejskal F., Zhu G., Keithly J.S., Martin W.;
RT "Pyruvate: NADP oxidoreductase from the mitochondrion of Euglena gracilis
RT and from the apicomplexan Cryptosporidium parvum: a biochemical relic
RT linking pyruvate metabolism in mitochondriate and amitochondriate
RT protists.";
RL Mol. Biol. Evol. 18:710-720(2001).
RN [3]
RP PROTEIN SEQUENCE OF 38-53 AND 1240-1259.
RX PubMed=1910287; DOI=10.1016/0003-9861(91)90040-p;
RA Inui H., Yamaji R., Saidoh H., Miyatake K., Nakano Y., Kitaoka S.;
RT "Pyruvate:NADP+ oxidoreductase from Euglena gracilis: limited proteolysis
RT of the enzyme with trypsin.";
RL Arch. Biochem. Biophys. 286:270-276(1991).
RN [4]
RP PROTEIN SEQUENCE OF 1240-1255, FUNCTION, AND CHARACTERIZATION.
RX PubMed=8373179; DOI=10.1006/abbi.1993.1441;
RA Watanabe F., Yamaji R., Isegawa Y., Yamamoto T., Tamura Y., Nakano Y.;
RT "Characterization of aquacobalamin reductase (NADPH) from Euglena
RT gracilis.";
RL Arch. Biochem. Biophys. 305:421-427(1993).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, SUBUNIT, COFACTOR, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBSTRATE SPECIFICITY.
RX PubMed=3110154; DOI=10.1016/s0021-9258(18)48057-x;
RA Inui H., Ono K., Miyatake K., Nakano Y., Kitaoka S.;
RT "Purification and characterization of pyruvate:NADP+ oxidoreductase in
RT Euglena gracilis.";
RL J. Biol. Chem. 262:9130-9135(1987).
RN [6]
RP FUNCTION, COFACTOR, SUBUNIT, AND SUBCELLULAR LOCATION.
RX PubMed=12623066; DOI=10.1016/s0003-9861(02)00749-x;
RA Nakazawa M., Takenaka S., Ueda M., Inui H., Nakano Y., Miyatake K.;
RT "Pyruvate:NADP(+) oxidoreductase is stabilized by its cofactor, thiamin
RT pyrophosphate, in mitochondria of Euglena gracilis.";
RL Arch. Biochem. Biophys. 411:183-188(2003).
CC -!- FUNCTION: Pyruvate dehydrogenase [NADP(+)] is one of three enzymes
CC participating in respiratory metabolism. The enzyme is also active with
CC 2-oxobutyrate and oxaloacetate. The enzyme is oxygen sensitive.
CC {ECO:0000269|PubMed:12623066, ECO:0000269|PubMed:3110154}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=CoA + NADP(+) + pyruvate = acetyl-CoA + CO2 + NADPH;
CC Xref=Rhea:RHEA:17425, ChEBI:CHEBI:15361, ChEBI:CHEBI:16526,
CC ChEBI:CHEBI:57287, ChEBI:CHEBI:57288, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.2.1.51;
CC Evidence={ECO:0000269|PubMed:3110154};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:12623066};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:12623066};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000269|PubMed:12623066};
CC Note=Binds 1 FMN per subunit. {ECO:0000269|PubMed:12623066};
CC -!- COFACTOR:
CC Name=thiamine diphosphate; Xref=ChEBI:CHEBI:58937;
CC Evidence={ECO:0000269|PubMed:12623066};
CC -!- COFACTOR:
CC Name=iron-sulfur cluster; Xref=ChEBI:CHEBI:30408;
CC Evidence={ECO:0000269|PubMed:3110154};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=210 uM for pyruvate {ECO:0000269|PubMed:3110154};
CC KM=8 uM for CoA {ECO:0000269|PubMed:3110154};
CC KM=30 uM for NADP(+) {ECO:0000269|PubMed:3110154};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:3110154};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:3110154};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:12623066,
CC ECO:0000269|PubMed:3110154}.
CC -!- SUBCELLULAR LOCATION: Mitochondrion {ECO:0000269|PubMed:11319255,
CC ECO:0000269|PubMed:12623066}.
CC -!- INDUCTION: Expressed in both aerobic and anaerobic conditions.
CC -!- MISCELLANEOUS: Arose from gene fusion of pyruvate:ferredoxin
CC oxidoreductase and cytochrome-P450 reductase. Gene fusion has only been
CC found in Euglena and Cryptosporidium. {ECO:0000303|PubMed:11023353,
CC ECO:0000303|PubMed:11319255}.
CC -!- MISCELLANEOUS: Euglena cells absolutely require thiamine for growth due
CC to the lack of a pyrimidine formation biosynthetic pathway. Thiamine is
CC actively taken up into the cells and is used as a cofactor after being
CC converted to TPP (thiamine pyrophosphate).
CC {ECO:0000269|PubMed:12623066}.
CC -!- SIMILARITY: In the N-terminal section; belongs to the
CC pyruvate:ferredoxin/flavodoxin oxidoreductase family. {ECO:0000305}.
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DR EMBL; AB021127; BAB12024.1; -; mRNA.
DR EMBL; AJ278425; CAC37628.1; -; mRNA.
DR PIR; S36876; S36876.
DR AlphaFoldDB; Q94IN5; -.
DR SMR; Q94IN5; -.
DR KEGG; ag:BAB12024; -.
DR BioCyc; MetaCyc:MONOMER-17049; -.
DR BRENDA; 1.2.1.51; 2197.
DR SABIO-RK; Q94IN5; -.
DR GO; GO:0005739; C:mitochondrion; IDA:UniProtKB.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IDA:UniProtKB.
DR GO; GO:0050243; F:pyruvate dehydrogenase (NADP+) activity; IEA:UniProtKB-EC.
DR GO; GO:0045333; P:cellular respiration; IDA:UniProtKB.
DR GO; GO:0022900; P:electron transport chain; IEA:InterPro.
DR GO; GO:0006090; P:pyruvate metabolic process; IDA:UniProtKB.
DR CDD; cd06207; CyPoR_like; 1.
DR CDD; cd03377; TPP_PFOR_PNO; 1.
DR CDD; cd07034; TPP_PYR_PFOR_IOR-alpha_like; 1.
DR Gene3D; 3.30.70.20; -; 1.
DR Gene3D; 3.40.50.360; -; 1.
DR Gene3D; 3.40.50.920; -; 1.
DR Gene3D; 3.40.50.970; -; 2.
DR Gene3D; 3.40.50.80; Nucleotide-binding domain of ferredoxin-NADP reductase (FNR) module; 1.
DR Gene3D; 3.40.920.10; Pyruvate-ferredoxin oxidoreductase, PFOR, domain III; 1.
DR Gene3D; 4.10.780.10; Pyruvate-flavodoxin oxidoreductase, EKR domain; 1.
DR Gene3D; 2.40.30.10; Translation factors; 1.
DR InterPro; IPR017896; 4Fe4S_Fe-S-bd.
DR InterPro; IPR017900; 4Fe4S_Fe_S_CS.
DR InterPro; IPR003097; CysJ-like_FAD-binding.
DR InterPro; IPR017927; FAD-bd_FR_type.
DR InterPro; IPR001094; Flavdoxin-like.
DR InterPro; IPR008254; Flavodoxin/NO_synth.
DR InterPro; IPR001709; Flavoprot_Pyr_Nucl_cyt_Rdtase.
DR InterPro; IPR029039; Flavoprotein-like_sf.
DR InterPro; IPR039261; FNR_nucleotide-bd.
DR InterPro; IPR023173; NADPH_Cyt_P450_Rdtase_alpha.
DR InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR InterPro; IPR033412; PFOR_II.
DR InterPro; IPR037112; Pyrv-flavodox_OxR_EKR_sf.
DR InterPro; IPR019456; Pyrv-flavodox_OxRtase_EKR.
DR InterPro; IPR019752; Pyrv/ketoisovalerate_OxRed_cat.
DR InterPro; IPR002880; Pyrv_Fd/Flavodoxin_OxRdtase_N.
DR InterPro; IPR011895; Pyrv_flavodox_OxRed.
DR InterPro; IPR002869; Pyrv_flavodox_OxRed_cen.
DR InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR InterPro; IPR029061; THDP-binding.
DR InterPro; IPR009014; Transketo_C/PFOR_II.
DR NCBIfam; TIGR02176; pyruv_ox_red; 1.
DR PANTHER; PTHR32154; PYRUVATE-FLAVODOXIN OXIDOREDUCTASE-RELATED; 1.
DR PANTHER; PTHR32154:SF0; PYRUVATE-FLAVODOXIN OXIDOREDUCTASE-RELATED; 1.
DR Pfam; PF10371; EKR; 1.
DR Pfam; PF00667; FAD_binding_1; 1.
DR Pfam; PF12838; Fer4_7; 1.
DR Pfam; PF00258; Flavodoxin_1; 1.
DR Pfam; PF00175; NAD_binding_1; 1.
DR Pfam; PF17147; PFOR_II; 1.
DR Pfam; PF01558; POR; 1.
DR Pfam; PF01855; POR_N; 1.
DR PRINTS; PR00369; FLAVODOXIN.
DR PRINTS; PR00371; FPNCR.
DR SMART; SM00890; EKR; 1.
DR SUPFAM; SSF54862; 4Fe-4S ferredoxins; 1.
DR SUPFAM; SSF52343; Ferredoxin reductase-like, C-terminal NADP-linked domain; 1.
DR SUPFAM; SSF52218; Flavoproteins; 1.
DR SUPFAM; SSF53323; Pyruvate-ferredoxin oxidoreductase, PFOR, domain III; 1.
DR SUPFAM; SSF63380; Riboflavin synthase domain-like; 1.
DR SUPFAM; SSF52518; Thiamin diphosphate-binding fold (THDP-binding); 2.
DR SUPFAM; SSF52922; TK C-terminal domain-like; 1.
DR PROSITE; PS00198; 4FE4S_FER_1; 2.
DR PROSITE; PS51379; 4FE4S_FER_2; 2.
DR PROSITE; PS51384; FAD_FR; 1.
DR PROSITE; PS50902; FLAVODOXIN_LIKE; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Direct protein sequencing; Electron transport; FAD; Flavoprotein;
KW FMN; Iron; Iron-sulfur; Metal-binding; Mitochondrion; NADP; Oxidoreductase;
KW Repeat; Thiamine pyrophosphate; Transit peptide; Transport.
FT TRANSIT 1..37
FT /note="Mitochondrion"
FT /evidence="ECO:0000269|PubMed:1910287"
FT CHAIN 38..1803
FT /note="Pyruvate dehydrogenase [NADP(+)], mitochondrial"
FT /id="PRO_0000023867"
FT DOMAIN 747..776
FT /note="4Fe-4S ferredoxin-type 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00711"
FT DOMAIN 802..831
FT /note="4Fe-4S ferredoxin-type 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00711"
FT DOMAIN 1248..1391
FT /note="Flavodoxin-like"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT DOMAIN 1425..1650
FT /note="FAD-binding FR-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT BINDING 756
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /evidence="ECO:0000255"
FT BINDING 759
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /evidence="ECO:0000255"
FT BINDING 762
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /evidence="ECO:0000255"
FT BINDING 766
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /evidence="ECO:0000255"
FT BINDING 811
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000255"
FT BINDING 814
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000255"
FT BINDING 817
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000255"
FT BINDING 821
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000255"
FT BINDING 1458..1469
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 1585..1595
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT CONFLICT 50
FT /note="P -> S (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 52
FT /note="H -> A (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 53
FT /note="V -> K (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 598..599
FT /note="NV -> KL (in Ref. 1; BAB12024)"
FT /evidence="ECO:0000305"
FT CONFLICT 1256
FT /note="T -> E (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 1558
FT /note="M -> T (in Ref. 1; BAB12024)"
FT /evidence="ECO:0000305"
FT CONFLICT 1673..1674
FT /note="ER -> DG (in Ref. 1; BAB12024)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 1803 AA; 199821 MW; 5AFB6E3869CADCC6 CRC64;
MKQSVRPIIS NVLRKEVALY STIIGQDKGK EPTGRTYTSG PKPASHIEVP HHVTVPATDR
TPNPDAQFFQ SVDGSQATSH VAYALSDTAF IYPITPSSVM GELADVWMAQ GRKNAFGQVV
DVREMQSEAG AAGALHGALA AGAIATTFTA SQGLLLMIPN MYKIAGELMP SVIHVAAREL
AGHALSIFGG HADVMAVRQT GWAMLCSHTV QQSHDMALIS HVATLKSSIP FVHFFDGFRT
SHEVNKIKML PYAELKKLVP PGTMEQHWAR SLNPMHPTIR GTNQSADIYF QNMESANQYY
TDLAEVVQET MDEVAPYIGR HYKIFEYVGA PDAEEVTVLM GSGATTVNEA VDLLVKRGKK
VGAVLVHLYR PWSTKAFEKV LPKTVKRIAA LDRCKEVTAL GEPLYLDVSA TLNLFPERQN
VKVIGGRYGL GSKDFIPEHA LAIYANLASE NPIQRFTVGI TDDVTGTSVP FVNERVDTLP
EGTRQCVFWG IGSDGTVGAN RSAVRIIGDN SDLMVQAYFQ FDAFKSGGVT SSHLRFGPKP
ITAQYLVTNA DYIACHFQEY VKRFDMLDAI REGGTFVLNS RWTTEDMEKE IPADFRRNVA
QKKVRFYNVD ARKICDSFGL GKRINMLMQA CFFKLSGVLP LAEAQRLLNE SIVHEYGKKG
GKVVEMNQAV VNAVFAGDLP QEVQVPAAWA NAVDTSTRTP TGIEFVDKIM RPLMDFKGDQ
LPVSVMTPGG TFPVGTTQYA KRAIAAFIPQ WIPANCTQCN YCSYVCPHAT IRPFVLTDQE
VQLAPESFVT RKAKGDYQGM NFRIQVAPED CTGCQVCVET CPDDALEMTD AFTATPVQRT
NWEFAIKVPN RGTMTDRYSL KGSQFQQPLL EFSGACEGCG ETPYVKLLTQ LFGERTVIAN
ATGCSSIWGG TAGLAPYTTN AKGQGPAWGN SLFEDNAEFG FGIAVANAQK RSRVRDCILQ
AVEKKVADEG LTTLLAQWLQ DWNTGDKTLK YQDQIIAGLA QQRSKDPLLE QIYGMKDMLP
NISQWIIGGD GWANDIGFGG LDHVLASGQN LNVLVLDTEM YSNTGGQASK STHMASVAKF
ALGGKRTNKK NLTEMAMSYG NVYVATVSHG NMAQCVKAFV EAESYDGPSL IVGYAPCIEH
GLRAGMARMV QESEAAIATG YWPLYRFDPR LATEGKNPFQ LDSKRIKGNL QEYLDRQNRY
VNLKKNNPKG ADLLKSQMAD NITARFNRYR RMLEGPNTKA AAPSGNHVTI LYGSETGNSE
GLAKELATDF ERREYSVAVQ ALDDIDVADL ENMGFVVIAV STCGQGQFPR NSQLFWRELQ
RDKPEGWLKN LKYTVFGLGD STYYFYCHTA KQIDARLAAL GAQRVVPIGF GDDGDEDMFH
TGFNNWIPSV WNELKTKTPE EALFTPSIAV QLTPNATPQD FHFAKSTPVL SITGAERITP
ADHTRNFVTI RWKTDLSYQV GDSLGVFPEN TRSVVEEFLQ YYGLNPKDVI TIENKGSREL
PHCMAVGDLF TKVLDILGKP NNRFYKTLSY FAVDKAEKER LLKIAEMGPE YSNILSEMYH
YADIFHMFPS ARPTLQYLIE MIPNIKPRYY SISSAPIHTP GEVHSLVLID TWITLSGKHR
TGLTCTMLEH LQAGQVVDGC IHPTAMEFPD HEKPVVMCAM GSGLAPFVAF LRERSTLRKQ
GKKTGNMALY FGNRYEKTEF LMKEELKGHI NDGLLTLRCA FSRDDPKKKV YVQDLIKMDE
KMMYDYLVVQ KGSMYCCGSR SFIKPVQESL KHCFMKAGGL TAEQAENEVI DMFTTGRYNI
EAW
//
