NROR_PYRFU
ID NROR_PYRFU Reviewed; 359 AA.
AC Q8U1K9;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2002, sequence version 1.
DT 03-AUG-2022, entry version 129.
DE RecName: Full=NAD(P)H:rubredoxin oxidoreductase {ECO:0000303|PubMed:10464233};
DE Short=NROR {ECO:0000303|PubMed:10464233};
DE EC=1.18.1.4 {ECO:0000269|PubMed:10464233, ECO:0000269|PubMed:11398485, ECO:0000269|PubMed:15746356};
DE AltName: Full=Rubredoxin--NAD(P)(+) reductase {ECO:0000305};
GN OrderedLocusNames=PF1197 {ECO:0000312|EMBL:AAL81321.1};
OS Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1).
OC Archaea; Euryarchaeota; Thermococci; Thermococcales; Thermococcaceae;
OC Pyrococcus.
OX NCBI_TaxID=186497;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 43587 / DSM 3638 / JCM 8422 / Vc1;
RX PubMed=10430560; DOI=10.1093/genetics/152.4.1299;
RA Maeder D.L., Weiss R.B., Dunn D.M., Cherry J.L., Gonzalez J.M.,
RA DiRuggiero J., Robb F.T.;
RT "Divergence of the hyperthermophilic archaea Pyrococcus furiosus and P.
RT horikoshii inferred from complete genomic sequences.";
RL Genetics 152:1299-1305(1999).
RN [2]
RP PROTEIN SEQUENCE OF 1-25, FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND SUBCELLULAR LOCATION.
RC STRAIN=ATCC 43587 / DSM 3638 / JCM 8422 / Vc1;
RX PubMed=10464233; DOI=10.1128/jb.181.17.5530-5533.1999;
RA Ma K., Adams M.W.;
RT "A hyperactive NAD(P)H:Rubredoxin oxidoreductase from the hyperthermophilic
RT archaeon Pyrococcus furiosus.";
RL J. Bacteriol. 181:5530-5533(1999).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBUNIT, AND SUBCELLULAR LOCATION.
RC STRAIN=ATCC 43587 / DSM 3638 / JCM 8422 / Vc1;
RX PubMed=11398485; DOI=10.1016/s0076-6879(01)34458-0;
RA Ma K., Adams M.W.;
RT "NAD(P)H:rubredoxin oxidoreductase from Pyrococcus furiosus.";
RL Methods Enzymol. 334:55-62(2001).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=ATCC 43587 / DSM 3638 / JCM 8422 / Vc1;
RX PubMed=15746356; DOI=10.1128/aem.71.3.1522-1530.2005;
RA Grunden A.M., Jenney F.E. Jr., Ma K., Ji M., Weinberg M.V., Adams M.W.;
RT "In vitro reconstitution of an NADPH-dependent superoxide reduction pathway
RT from Pyrococcus furiosus.";
RL Appl. Environ. Microbiol. 71:1522-1530(2005).
RN [5] {ECO:0007744|PDB:1XHC}
RP X-RAY CRYSTALLOGRAPHY (2.35 ANGSTROMS) OF 2-359 IN COMPLEX WITH FAD, AND
RP COFACTOR.
RC STRAIN=ATCC 43587 / DSM 3638 / JCM 8422 / Vc1;
RG Southeast collaboratory for structural genomics (SECSG);
RA Horanyi P., Tempel W., Weinberg M.V., Liu Z.-J., Shah A., Chen L., Lee D.,
RA Sugar F.J., Brereton P.S., Izumi M., Poole F.L. II, Shah C.,
RA Jenney F.E. Jr., Arendall W.B. III, Rose J.P., Adams M.W.W.,
RA Richardson J.S., Richardson D.C., Wang B.-C.;
RT "NADH oxidase /nitrite reductase from Pyrococcus furiosus Pfu-1140779-
RT 001.";
RL Submitted (SEP-2004) to the PDB data bank.
CC -!- FUNCTION: Catalyzes the NADH-dependent reduction of rubredoxin (Rd), a
CC small iron-containing redox protein (PubMed:10464233, PubMed:11398485,
CC PubMed:15746356). Can also reduce other electron carriers, including
CC 2,6-dichlorophenol indophenol, benzyl viologen and menadione, but
CC rubredoxin is the most efficient electron acceptor (PubMed:10464233,
CC PubMed:11398485, PubMed:15746356). Does not reduce ferredoxin
CC (PubMed:10464233). Shows comparable activity with NADPH and NADH as
CC electron donors, but NADPH is probably the preferred physiological
CC electron donor (PubMed:10464233, PubMed:11398485). Is probably part of
CC an oxygen detoxification system that protects P.furiosus during
CC exposure to oxygen (PubMed:15746356). {ECO:0000269|PubMed:10464233,
CC ECO:0000269|PubMed:11398485, ECO:0000269|PubMed:15746356}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + NADP(+) + 2 reduced [rubredoxin] = NADPH + 2 oxidized
CC [rubredoxin]; Xref=Rhea:RHEA:13949, Rhea:RHEA-COMP:10302, Rhea:RHEA-
CC COMP:10303, ChEBI:CHEBI:15378, ChEBI:CHEBI:29033, ChEBI:CHEBI:29034,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.18.1.4;
CC Evidence={ECO:0000269|PubMed:10464233, ECO:0000269|PubMed:11398485,
CC ECO:0000269|PubMed:15746356};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + NAD(+) + 2 reduced [rubredoxin] = NADH + 2 oxidized
CC [rubredoxin]; Xref=Rhea:RHEA:18597, Rhea:RHEA-COMP:10302, Rhea:RHEA-
CC COMP:10303, ChEBI:CHEBI:15378, ChEBI:CHEBI:29033, ChEBI:CHEBI:29034,
CC ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; EC=1.18.1.4;
CC Evidence={ECO:0000269|PubMed:10464233, ECO:0000269|PubMed:11398485,
CC ECO:0000269|PubMed:15746356};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:10464233, ECO:0000269|PubMed:11398485,
CC ECO:0000269|PubMed:15746356};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|Ref.5};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.05 mM for rubredoxin (at 80 degrees Celsius, in the presence of
CC NADPH) {ECO:0000269|PubMed:10464233};
CC KM=0.01 mM for rubredoxin (at 25 degrees Celsius, in the presence of
CC NADPH) {ECO:0000269|PubMed:10464233};
CC KM=0.037 mM for rubredoxin (for recombinant enzyme, at 80 degrees
CC Celsius, in the presence of NADPH) {ECO:0000269|PubMed:15746356};
CC KM=0.0016 mM for rubredoxin (for recombinant enzyme, at 23 degrees
CC Celsius, in the presence of NADPH) {ECO:0000269|PubMed:15746356};
CC KM=1.25 mM for benzyl viologen (at 80 degrees Celsius, in the
CC presence of NADH) {ECO:0000269|PubMed:10464233};
CC KM=5.5 mM for benzyl viologen (for recombinant enzyme, at 80 degrees
CC Celsius, in the presence of NADH) {ECO:0000269|PubMed:15746356};
CC KM=0.25 mM for 5,5'-dithiobis[2-nitrobenzoic acid] (at 80 degrees
CC Celsius, in the presence of NADPH) {ECO:0000269|PubMed:10464233};
CC KM=9.5 mM for H(2)O(2) (at 80 degrees Celsius, in the presence of
CC NADPH) {ECO:0000269|PubMed:10464233};
CC KM=0.005 mM for NADPH (at 80 degrees Celsius, in the presence of
CC benzyl viologen) {ECO:0000269|PubMed:10464233};
CC KM=0.048 mM for NADPH (for recombinant enzyme, at 80 degrees Celsius,
CC in the presence of benzyl viologen) {ECO:0000269|PubMed:15746356};
CC KM=0.034 mM for NADH (at 80 degrees Celsius, in the presence of
CC benzyl viologen) {ECO:0000269|PubMed:10464233};
CC KM=0.18 mM for NADH (for recombinant enzyme, at 80 degrees Celsius,
CC in the presence of benzyl viologen) {ECO:0000269|PubMed:15746356};
CC Vmax=20000 umol/min/mg enzyme with rubredoxin as substrate (at 80
CC degrees Celsius, in the presence of NADPH)
CC {ECO:0000269|PubMed:10464233};
CC Vmax=294 umol/min/mg enzyme with rubredoxin as substrate (at 25
CC degrees Celsius, in the presence of NADPH)
CC {ECO:0000269|PubMed:10464233};
CC Vmax=7307 umol/min/mg enzyme with rubredoxin as substrate (for
CC recombinant enzyme, at 80 degrees Celsius, in the presence of NADPH)
CC {ECO:0000269|PubMed:15746356};
CC Vmax=526 umol/min/mg enzyme with rubredoxin as substrate (for
CC recombinant enzyme, at 23 degrees Celsius, in the presence of NADPH)
CC {ECO:0000269|PubMed:15746356};
CC Vmax=1000 umol/min/mg enzyme with benzyl viologen as substrate (at 80
CC degrees Celsius, in the presence of NADH)
CC {ECO:0000269|PubMed:10464233};
CC Vmax=1000 umol/min/mg enzyme with benzyl viologen as substrate (for
CC recombinant enzyme, at 80 degrees Celsius, in the presence of NADH)
CC {ECO:0000269|PubMed:15746356};
CC Vmax=25 umol/min/mg enzyme with 5,5'-dithiobis[2-nitrobenzoic acid]
CC as substrate (at 80 degrees Celsius, in the presence of NADPH)
CC {ECO:0000269|PubMed:10464233};
CC Vmax=12 umol/min/mg enzyme with H(2)O(2) as substrate (at 80 degrees
CC Celsius, in the presence of NADPH) {ECO:0000269|PubMed:10464233};
CC Vmax=455 umol/min/mg enzyme with NADPH as substrate (at 80 degrees
CC Celsius, in the presence of benzyl viologen)
CC {ECO:0000269|PubMed:10464233};
CC Vmax=208 umol/min/mg enzyme with NADPH as substrate (for recombinant
CC enzyme, at 80 degrees Celsius, in the presence of benzyl viologen)
CC {ECO:0000269|PubMed:15746356};
CC Vmax=472 umol/min/mg enzyme with NADH as substrate (at 80 degrees
CC Celsius, in the presence of benzyl viologen)
CC {ECO:0000269|PubMed:10464233};
CC Vmax=400 umol/min/mg enzyme with NADH as substrate (for recombinant
CC enzyme, at 80 degrees Celsius, in the presence of benzyl viologen)
CC {ECO:0000269|PubMed:15746356};
CC Note=kcat is 15000 sec(-1) with rubredoxin as substrate. kcat is 750
CC sec(-1) with benzyl viologen as substrate. kcat is 19 sec(-1) with
CC 5,5'-dithiobis[2-nitrobenzoic acid] as substrate. kcat is 9 sec(-1)
CC with H(2)O(2) as substrate. kcat is 341 sec(-1) with NADPH as
CC substrate. kcat is 354 sec(-1) with NADH as substrate.
CC {ECO:0000269|PubMed:10464233};
CC pH dependence:
CC Optimum pH is 7.0 for the reduction of rubredoxin.
CC {ECO:0000269|PubMed:10464233};
CC Temperature dependence:
CC Thermostable. {ECO:0000269|PubMed:10464233};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:10464233,
CC ECO:0000269|PubMed:11398485}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:10464233,
CC ECO:0000269|PubMed:11398485}.
CC -!- MISCELLANEOUS: NROR, unlike the vast majority of enzymes from
CC hyperthermophilic sources, shows considerable activity at low
CC temperatures. {ECO:0000269|PubMed:10464233,
CC ECO:0000269|PubMed:15746356}.
CC -!- SIMILARITY: Belongs to the FAD-dependent oxidoreductase family.
CC {ECO:0000305}.
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DR EMBL; AE009950; AAL81321.1; -; Genomic_DNA.
DR RefSeq; WP_011012338.1; NZ_CP023154.1.
DR PDB; 1XHC; X-ray; 2.35 A; A=2-359.
DR PDBsum; 1XHC; -.
DR SMR; Q8U1K9; -.
DR STRING; 186497.PF1197; -.
DR PRIDE; Q8U1K9; -.
DR EnsemblBacteria; AAL81321; AAL81321; PF1197.
DR GeneID; 41713005; -.
DR KEGG; pfu:PF1197; -.
DR PATRIC; fig|186497.12.peg.1258; -.
DR eggNOG; arCOG01070; Archaea.
DR HOGENOM; CLU_003291_4_4_2; -.
DR OMA; IATYPYH; -.
DR OrthoDB; 33756at2157; -.
DR PhylomeDB; Q8U1K9; -.
DR BRENDA; 1.18.1.4; 5243.
DR EvolutionaryTrace; Q8U1K9; -.
DR Proteomes; UP000001013; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.30.390.30; -; 1.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR023753; FAD/NAD-binding_dom.
DR InterPro; IPR016156; FAD/NAD-linked_Rdtase_dimer_sf.
DR Pfam; PF07992; Pyr_redox_2; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR SUPFAM; SSF55424; SSF55424; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Direct protein sequencing; Electron transport;
KW FAD; Flavoprotein; NAD; NADP; Nucleotide-binding; Oxidoreductase;
KW Reference proteome; Stress response; Transport.
FT CHAIN 1..359
FT /note="NAD(P)H:rubredoxin oxidoreductase"
FT /id="PRO_0000454941"
FT BINDING 11
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT BINDING 29..30
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT BINDING 38..39
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT BINDING 75
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT BINDING 103
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT BINDING 120
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT BINDING 260
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.5, ECO:0007744|PDB:1XHC"
FT STRAND 2..6
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 10..19
FT /evidence="ECO:0007829|PDB:1XHC"
FT TURN 20..22
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 23..28
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 30..33
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 41..45
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 51..54
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 59..65
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 67..70
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 75..79
FT /evidence="ECO:0007829|PDB:1XHC"
FT TURN 80..83
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 84..89
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 91..93
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 95..99
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 103..105
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 113..115
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 122..135
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 136..142
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 145..156
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 160..164
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 166..168
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 175..187
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 190..193
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 198..201
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 203..208
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 211..214
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 218..221
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 224..226
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 229..233
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 238..243
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 255..257
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 259..261
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 262..264
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 272..286
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 299..304
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 307..313
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 319..323
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 326..329
FT /evidence="ECO:0007829|PDB:1XHC"
FT STRAND 337..342
FT /evidence="ECO:0007829|PDB:1XHC"
FT HELIX 344..350
FT /evidence="ECO:0007829|PDB:1XHC"
SQ SEQUENCE 359 AA; 39941 MW; 79891673B83E98C7 CRC64;
MKVVIVGNGP GGFELAKQLS QTYEVTVIDK EPVPYYSKPM LSHYIAGFIP RNRLFPYSLD
WYRKRGIEIR LAEEAKLIDR GRKVVITEKG EVPYDTLVLA TGARAREPQI KGKEYLLTLR
TIFDADRIKE SIENSGEAII IGGGFIGLEL AGNLAEAGYH VKLIHRGAMF LGLDEELSNM
IKDMLEETGV KFFLNSELLE ANEEGVLTNS GFIEGKVKIC AIGIVPNVDL ARRSGIHTGR
GILIDDNFRT SAKDVYAIGD CAEYSGIIAG TAKAAMEQAR VLADILKGEP RRYNFKFRST
VFKFGKLQIA IIGNTKGEGK WIEDNTKVFY ENGKIIGAVV FNDIRKATKL EKEILDFYS
