HMP_CUPNH
ID HMP_CUPNH Reviewed; 403 AA.
AC P39662; Q7WXD4;
DT 01-FEB-1995, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 2.
DT 03-AUG-2022, entry version 169.
DE RecName: Full=Flavohemoprotein;
DE AltName: Full=FHP;
DE AltName: Full=Flavohemoglobin;
DE AltName: Full=Hemoglobin-like protein;
DE AltName: Full=Nitric oxide dioxygenase;
DE Short=NO oxygenase;
DE Short=NOD;
DE EC=1.14.12.17;
GN Name=hmp; Synonyms=fhp; OrderedLocusNames=PHG200;
OS Cupriavidus necator (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442
OS / H16 / Stanier 337) (Ralstonia eutropha).
OG Plasmid megaplasmid pHG1.
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Burkholderiaceae; Cupriavidus.
OX NCBI_TaxID=381666;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=8125952; DOI=10.1016/s0021-9258(17)37291-5;
RA Cramm R., Siddiqui R.A., Friedrich B.;
RT "Primary sequence and evidence for a physiological function of the
RT flavohemoprotein of Alcaligenes eutrophus.";
RL J. Biol. Chem. 269:7349-7354(1994).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337;
RX PubMed=12948488; DOI=10.1016/s0022-2836(03)00894-5;
RA Schwartz E., Henne A., Cramm R., Eitinger T., Friedrich B., Gottschalk G.;
RT "Complete nucleotide sequence of pHG1: a Ralstonia eutropha H16 megaplasmid
RT encoding key enzymes of H(2)-based lithoautotrophy and anaerobiosis.";
RL J. Mol. Biol. 332:369-383(2003).
RN [3]
RP PARTIAL PROTEIN SEQUENCE.
RX PubMed=1594608; DOI=10.1073/pnas.89.11.5015;
RA Zhu H., Riggs A.F.;
RT "Yeast flavohemoglobin is an ancient protein related to globins and a
RT reductase family.";
RL Proc. Natl. Acad. Sci. U.S.A. 89:5015-5019(1992).
RN [4]
RP COFACTOR, INDUCTION, AND SPECTRAL PROPERTIES.
RX PubMed=218634; DOI=10.1016/0005-2795(79)90422-7;
RA Probst I., Wolf G., Schlegel H.G.;
RT "An oxygen-binding flavohemoprotein from Alcaligenes eutrophus.";
RL Biochim. Biophys. Acta 576:471-478(1979).
RN [5]
RP ENZYME ACTIVITY.
RX PubMed=10922365; DOI=10.1074/jbc.m004141200;
RA Gardner P.R., Gardner A.M., Martin L.A., Dou Y., Li T., Olson J.S., Zhu H.,
RA Riggs A.F.;
RT "Nitric-oxide dioxygenase activity and function of flavohemoglobins.
RT Sensitivity to nitric oxide and carbon monoxide inhibition.";
RL J. Biol. Chem. 275:31581-31587(2000).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS).
RX PubMed=8557026; DOI=10.1002/j.1460-2075.1995.tb00297.x;
RA Ermler U., Siddiqui R.A., Cramm R., Friedrich B.;
RT "Crystal structure of the flavohemoglobin from Alcaligenes eutrophus at
RT 1.75-A resolution.";
RL EMBO J. 14:6067-6077(1995).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF NATIVE PROTEIN AND MUTANT V98F,
RP PHOSPHOLIPID BINDING, AND MUTAGENESIS OF ALA-60 AND VAL-98.
RX PubMed=10336624; DOI=10.1046/j.1432-1327.1999.00381.x;
RA Ollesch G., Kaunzinger A., Juchelka D., Schubert-Zsilavecz M., Ermler U.;
RT "Phospholipid bound to the flavohemoprotein from Alcaligenes eutrophus.";
RL Eur. J. Biochem. 262:396-405(1999).
CC -!- FUNCTION: Is involved in NO detoxification in an aerobic process,
CC termed nitric oxide dioxygenase (NOD) reaction that utilizes O(2) and
CC NAD(P)H to convert NO to nitrate, which protects the bacterium from
CC various noxious nitrogen compounds. Therefore, plays a central role in
CC the inducible response to nitrosative stress.
CC -!- FUNCTION: In the presence of oxygen and NADH, FHP has NADH oxidase
CC activity, which leads to the generation of superoxide and H(2)O(2),
CC both in vitro and in vivo, and it has been suggested that FHP might act
CC as an amplifier of superoxide stress. Under anaerobic conditions, FHP
CC also exhibits nitric oxide reductase and FAD reductase activities.
CC However, all these reactions are much lower than NOD activity.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NADPH + 2 nitric oxide + 2 O2 = H(+) + NADP(+) + 2 nitrate;
CC Xref=Rhea:RHEA:19465, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16480, ChEBI:CHEBI:17632, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.14.12.17;
CC Evidence={ECO:0000269|PubMed:10922365};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NADH + 2 nitric oxide + 2 O2 = H(+) + NAD(+) + 2 nitrate;
CC Xref=Rhea:RHEA:19469, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16480, ChEBI:CHEBI:17632, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945; EC=1.14.12.17;
CC Evidence={ECO:0000269|PubMed:10922365};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692; Evidence={ECO:0000269|PubMed:218634};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:218634};
CC -!- COFACTOR:
CC Name=heme b; Xref=ChEBI:CHEBI:60344;
CC Evidence={ECO:0000269|PubMed:218634};
CC Note=Binds 1 heme b group per subunit. {ECO:0000269|PubMed:218634};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.10 uM for NO;
CC KM=80 uM for O(2);
CC KM=70 uM for NADH;
CC -!- SUBUNIT: Monomer.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- INDUCTION: Under oxygen-limited conditions.
CC {ECO:0000269|PubMed:218634}.
CC -!- DOMAIN: Consists of two distinct domains; an N-terminal heme-containing
CC oxygen-binding domain and a C-terminal reductase domain with binding
CC sites for FAD and NAD(P)H.
CC -!- MISCELLANEOUS: No protein-heme interactions have been detected at the
CC distal side of the heme molecule.
CC -!- MISCELLANEOUS: FHP is able to bind phospholipids with high affinity.
CC -!- SIMILARITY: Belongs to the globin family. Two-domain flavohemoproteins
CC subfamily. {ECO:0000305}.
CC -!- SIMILARITY: In the C-terminal section; belongs to the flavoprotein
CC pyridine nucleotide cytochrome reductase family. {ECO:0000305}.
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DR EMBL; X74334; CAA52381.1; -; Genomic_DNA.
DR EMBL; AY305378; AAP85952.1; -; Genomic_DNA.
DR PIR; A53396; A53396.
DR RefSeq; WP_011154115.1; NZ_CP039289.1.
DR PDB; 1CQX; X-ray; 1.75 A; A/B=1-403.
DR PDB; 3OZU; X-ray; 2.00 A; A=1-403.
DR PDB; 3OZV; X-ray; 2.40 A; A/B=1-403.
DR PDB; 3OZW; X-ray; 2.30 A; A/B=1-403.
DR PDBsum; 1CQX; -.
DR PDBsum; 3OZU; -.
DR PDBsum; 3OZV; -.
DR PDBsum; 3OZW; -.
DR AlphaFoldDB; P39662; -.
DR SMR; P39662; -.
DR STRING; 381666.PHG200; -.
DR DrugBank; DB03979; 1-[Glycerolylphosphonyl]-2-[8-(2-Hexyl-Cyclopropyl)-Octanal-1-Yl]-3-[Hexadecanal-1-Yl]-Glycerol.
DR DrugBank; DB03147; Flavin adenine dinucleotide.
DR EnsemblBacteria; AAP85952; AAP85952; PHG200.
DR GeneID; 39976365; -.
DR KEGG; reh:PHG200; -.
DR PATRIC; fig|381666.6.peg.150; -.
DR eggNOG; COG1017; Bacteria.
DR eggNOG; COG1018; Bacteria.
DR HOGENOM; CLU_003827_12_0_4; -.
DR OMA; EENHTEF; -.
DR OrthoDB; 1834577at2; -.
DR BRENDA; 1.14.12.17; 231.
DR SABIO-RK; P39662; -.
DR EvolutionaryTrace; P39662; -.
DR Proteomes; UP000008210; Plasmid megaplasmid pHG1.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008941; F:nitric oxide dioxygenase activity; IDA:CACAO.
DR GO; GO:0019825; F:oxygen binding; IEA:InterPro.
DR GO; GO:0005344; F:oxygen carrier activity; IEA:UniProtKB-UniRule.
DR GO; GO:0051409; P:response to nitrosative stress; IEA:InterPro.
DR GO; GO:0009636; P:response to toxic substance; IEA:UniProtKB-KW.
DR Gene3D; 1.10.490.10; -; 1.
DR Gene3D; 3.40.50.80; -; 1.
DR HAMAP; MF_01252; Hmp; 1.
DR InterPro; IPR008333; Cbr1-like_FAD-bd_dom.
DR InterPro; IPR017927; FAD-bd_FR_type.
DR InterPro; IPR039261; FNR_nucleotide-bd.
DR InterPro; IPR000971; Globin.
DR InterPro; IPR009050; Globin-like_sf.
DR InterPro; IPR012292; Globin/Proto.
DR InterPro; IPR023950; Hmp.
DR InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR PANTHER; PTHR43396:SF3; PTHR43396:SF3; 1.
DR Pfam; PF00970; FAD_binding_6; 1.
DR Pfam; PF00042; Globin; 1.
DR Pfam; PF00175; NAD_binding_1; 1.
DR SUPFAM; SSF46458; SSF46458; 1.
DR SUPFAM; SSF52343; SSF52343; 1.
DR SUPFAM; SSF63380; SSF63380; 1.
DR PROSITE; PS51384; FAD_FR; 1.
DR PROSITE; PS01033; GLOBIN; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Detoxification; Direct protein sequencing; FAD;
KW Flavoprotein; Heme; Iron; Metal-binding; NAD; NADP; Oxidoreductase;
KW Oxygen transport; Plasmid; Reference proteome; Transport.
FT CHAIN 1..403
FT /note="Flavohemoprotein"
FT /id="PRO_0000052441"
FT DOMAIN 152..262
FT /note="FAD-binding FR-type"
FT REGION 1..140
FT /note="Globin"
FT REGION 149..403
FT /note="Reductase"
FT ACT_SITE 95
FT /note="Charge relay system"
FT ACT_SITE 137
FT /note="Charge relay system"
FT BINDING 85
FT /ligand="heme b"
FT /ligand_id="ChEBI:CHEBI:60344"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="proximal binding residue"
FT BINDING 190
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT BINDING 206..209
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT BINDING 275..280
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250"
FT BINDING 395..398
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT SITE 29
FT /note="Involved in heme-bound ligand stabilization and O-O
FT bond activation"
FT SITE 84
FT /note="Influences the redox potential of the prosthetic
FT heme and FAD groups"
FT SITE 394
FT /note="Influences the redox potential of the prosthetic
FT heme and FAD groups"
FT MUTAGEN 60
FT /note="A->Y: Does not affect phospholipid-binding."
FT /evidence="ECO:0000269|PubMed:10336624"
FT MUTAGEN 98
FT /note="V->F: Blocks phospholipid-binding."
FT /evidence="ECO:0000269|PubMed:10336624"
FT CONFLICT 218
FT /note="S -> T (in Ref. 1; CAA52381)"
FT /evidence="ECO:0000305"
FT HELIX 4..19
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 21..35
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 37..41
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 47..49
FT /evidence="ECO:0007829|PDB:3OZW"
FT HELIX 50..67
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 71..88
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 92..94
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 95..110
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 111..113
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 116..145
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 155..164
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 166..176
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 190..196
FT /evidence="ECO:0007829|PDB:1CQX"
FT TURN 198..200
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 201..209
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 219..224
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 229..231
FT /evidence="ECO:0007829|PDB:3OZW"
FT HELIX 235..243
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 249..252
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 269..275
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 278..288
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 290..292
FT /evidence="ECO:0007829|PDB:3OZW"
FT STRAND 296..303
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 305..307
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 309..320
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 324..332
FT /evidence="ECO:0007829|PDB:1CQX"
FT TURN 339..341
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 344..348
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 351..353
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 355..358
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 364..370
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 371..383
FT /evidence="ECO:0007829|PDB:1CQX"
FT HELIX 388..390
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 391..393
FT /evidence="ECO:0007829|PDB:1CQX"
FT STRAND 396..398
FT /evidence="ECO:0007829|PDB:3OZV"
SQ SEQUENCE 403 AA; 44782 MW; 2E3ED365087F5EA0 CRC64;
MLTQKTKDIV KATAPVLAEH GYDIIKCFYQ RMFEAHPELK NVFNMAHQEQ GQQQQALARA
VYAYAENIED PNSLMAVLKN IANKHASLGV KPEQYPIVGE HLLAAIKEVL GNAATDDIIS
AWAQAYGNLA DVLMGMESEL YERSAEQPGG WKGWRTFVIR EKRPESDVIT SFILEPADGG
PVVNFEPGQY TSVAIDVPAL GLQQIRQYSL SDMPNGRSYR ISVKREGGGP QPPGYVSNLL
HDHVNVGDQV KLAAPYGSFH IDVDAKTPIV LISGGVGLTP MVSMLKVALQ APPRQVVFVH
GARNSAVHAM RDRLREAAKT YENLDLFVFY DQPLPEDVQG RDYDYPGLVD VKQIEKSILL
PDADYYICGP IPFMRMQHDA LKNLGIHEAR IHYEVFGPDL FAE
