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 27-MAR-2024, entry version 174. 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; Pseudomonadota; 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 RC 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=0.1 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}. 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; 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 DrugBank; DB03979; 1-[Glycerolylphosphonyl]-2-[8-(2-Hexyl-Cyclopropyl)-Octanal-1-Yl]-3-[Hexadecanal-1-Yl]-Glycerol. DR DrugBank; DB03147; Flavin adenine dinucleotide. 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 OrthoDB; 9801223at2; -. 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 CDD; cd14779; FHP_Ae-globin-like; 1. DR CDD; cd06184; flavohem_like_fad_nad_binding; 1. DR Gene3D; 1.10.490.10; Globins; 1. DR Gene3D; 3.40.50.80; Nucleotide-binding domain of ferredoxin-NADP reductase (FNR) module; 1. DR Gene3D; 2.40.30.10; Translation factors; 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; FLAVOHEMOPROTEIN; 1. DR PANTHER; PTHR43396:SF3; FLAVOHEMOPROTEIN; 1. DR Pfam; PF00970; FAD_binding_6; 1. DR Pfam; PF00042; Globin; 1. DR Pfam; PF00175; NAD_binding_1; 1. DR PRINTS; PR00410; PHEHYDRXLASE. DR SUPFAM; SSF52343; Ferredoxin reductase-like, C-terminal NADP-linked domain; 1. DR SUPFAM; SSF46458; Globin-like; 1. DR SUPFAM; SSF63380; Riboflavin synthase domain-like; 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 //