Reviewed, UniProtKB/Swiss-Prot P39662 (HMP_RALEH)

Names and origin

Protein names

Recommended name:
    Flavohemoprotein
Alternative name(s):
    Hemoglobin-like protein
    Flavohemoglobin
    FHP
    Nitric oxide dioxygenase
      Short name=NO oxygenase
      Short name=NOD
    EC=1.14.12.17

Gene names

Name:	hmp
Synonyms:	fhp
Ordered Locus Names:	PHG200

Encoded on

Plasmid megaplasmid pHG1

Organism

Ralstonia eutropha (strain ATCC 17699 / H16 / DSM 428 / Stanier 337) (Cupriavidus necator (strain ATCC 17699 / H16 / DSM 428 / Stanier 337)) [Complete proteome] [HAMAP]

Taxonomic identifier

381666 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Betaproteobacteria › Burkholderiales › Burkholderiaceae › Cupriavidus

Protein attributes

Sequence length	403 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Is involved in NO detoxification in an aerobic process, termed nitric oxide dioxygenase (NOD) reaction that utilizes O2 and NAD(P)H to convert NO to nitrate, which protects the bacterium from various noxious nitrogen compounds. Therefore, plays a central role in the inducible response to nitrosative stress. HAMAP MF_01252 In the presence of oxygen and NADH, FHP has NADH oxidase activity, which leads to the generation of superoxide and H2O2, both in vitro and in vivo, and it has been suggested that FHP might act as an amplifier of superoxide stress. Under anaerobic conditions, FHP also exhibits nitric oxide reductase and FAD reductase activities. However, all these reactions are much lower than NOD activity. HAMAP MF_01252
Catalytic activity	2 NO + 2 O2 + NAD(P)H = 2 NO3- + NAD(P)+. Ref.5
Cofactor	Binds 1 FAD per subunit. Ref.4 Binds 1 heme B group per subunit. Ref.4
Subunit structure	Monomer. HAMAP MF_01252
Subcellular location	Cytoplasm HAMAP MF_01252.
Induction	Under oxygen-limited conditions. Ref.4
Domain	Consists of two distinct domains; an N-terminal heme-containing oxygen-binding domain and a C-terminal reductase domain with binding sites for FAD and NAD(P)H. HAMAP MF_01252
Miscellaneous	No protein-heme interactions have been detected at the distal side of the heme molecule. HAMAP MF_01252 FHP is able to bind phospholipids with high affinity. HAMAP MF_01252
Sequence similarities	Belongs to the globin family. Two-domain flavohemoproteins subfamily. In the C-terminal section; belongs to the flavoprotein pyridine nucleotide cytochrome reductase family. Contains 1 FAD-binding FR-type domain.
Biophysicochemical properties	Kinetic parameters: KM=0.10 µM for NO HAMAP MF_01252 KM=80 µM for O2 KM=70 µM for NADH

Ontologies

Keywords
Biological process	Detoxification Oxygen transport Transport
Cellular component	Cytoplasm
Ligand	FAD Flavoprotein Heme Iron Metal-binding NAD NADP
Molecular function	Oxidoreductase
Technical term	3D-structure Complete proteome Direct protein sequencing Plasmid
Gene Ontology (GO)
Biological process	oxidation reduction Inferred from electronic annotation. Source: UniProtKB-KW oxygen transport Inferred from electronic annotation. Source: HAMAP response to toxin Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	heme binding Inferred from electronic annotation. Source: InterPro nitric oxide dioxygenase activity Inferred from electronic annotation. Source: HAMAP oxygen binding Inferred from electronic annotation. Source: InterPro oxygen transporter activity Inferred from electronic annotation. Source: HAMAP
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 403

403

Flavohemoprotein HAMAP MF_01252

PRO_0000052441

Regions

Domain

152 – 262

111

FAD-binding FR-type

Nucleotide binding

206 – 209

4

FAD HAMAP MF_01252

Nucleotide binding

275 – 280

6

NADP By similarity

Nucleotide binding

395 – 398

4

FAD HAMAP MF_01252

Region

1 – 140

140

Globin HAMAP MF_01252

Region

149 – 403

255

Reductase HAMAP MF_01252

Region

266 – 403

138

NAD or NADP-binding HAMAP MF_01252

Sites

Active site

95

1

Charge relay system HAMAP MF_01252

Active site

137

1

Charge relay system HAMAP MF_01252

Metal binding

85

1

Iron (heme proximal ligand) HAMAP MF_01252

Binding site

190

1

FAD HAMAP MF_01252

Site

29

1

Involved in heme-bound ligand stabilization and O-O bond activation HAMAP MF_01252

Site

84

1

Influences the redox potential of the prosthetic heme and FAD groups HAMAP MF_01252

Site

394

1

Influences the redox potential of the prosthetic heme and FAD groups HAMAP MF_01252

Experimental info

Mutagenesis

60

1

A → Y: Does not affect phospholipid-binding. Ref.7

Mutagenesis

98

1

V → F: Blocks phospholipid-binding. Ref.7

Sequence conflict

218

1

S → T in CAA52381. Ref.1

Secondary structure

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403

Helix Strand Turn

Details...

Helix

4 – 19

16

Helix

21 – 35

15

Helix

37 – 41

5

Helix

50 – 67

18

Helix

71 – 88

18

Helix

92 – 94

3

Helix

95 – 110

16

Helix

111 – 113

3

Helix

116 – 145

30

Beta strand

155 – 164

10

Beta strand

166 – 176

11

Beta strand

190 – 196

7

Turn

198 – 200

3

Beta strand

201 – 209

9

Beta strand

219 – 224

6

Helix

235 – 243

9

Beta strand

249 – 252

4

Beta strand

269 – 275

7

Helix

278 – 288

11

Beta strand

296 – 303

8

Beta strand

305 – 307

3

Helix

309 – 320

12

Beta strand

324 – 332

9

Turn

339 – 341

3

Beta strand

344 – 348

5

Helix

351 – 353

3

Helix

355 – 358

4

Beta strand

364 – 370

7

Helix

371 – 383

13

Helix

388 – 390

3

Beta strand

391 – 393

3

Sequences

	Sequence		Length	Mass (Da)
	P39662-1 [UniParc]. Last modified January 23, 2007. Version 2. Checksum: 2E3ED365087F5EA0	FASTA	403	44,782
	10 20 30 40 50 60 MLTQKTKDIV KATAPVLAEH GYDIIKCFYQ RMFEAHPELK NVFNMAHQEQ GQQQQALARA 70 80 90 100 110 120 VYAYAENIED PNSLMAVLKN IANKHASLGV KPEQYPIVGE HLLAAIKEVL GNAATDDIIS 130 140 150 160 170 180 AWAQAYGNLA DVLMGMESEL YERSAEQPGG WKGWRTFVIR EKRPESDVIT SFILEPADGG 190 200 210 220 230 240 PVVNFEPGQY TSVAIDVPAL GLQQIRQYSL SDMPNGRSYR ISVKREGGGP QPPGYVSNLL 250 260 270 280 290 300 HDHVNVGDQV KLAAPYGSFH IDVDAKTPIV LISGGVGLTP MVSMLKVALQ APPRQVVFVH 310 320 330 340 350 360 GARNSAVHAM RDRLREAAKT YENLDLFVFY DQPLPEDVQG RDYDYPGLVD VKQIEKSILL 370 380 390 400 PDADYYICGP IPFMRMQHDA LKNLGIHEAR IHYEVFGPDL FAE

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Primary sequence and evidence for a physiological function of the flavohemoprotein of Alcaligenes eutrophus." Cramm R., Siddiqui R.A., Friedrich B. J. Biol. Chem. 269:7349-7354(1994) [PubMed: 8125952] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"Complete nucleotide sequence of pHG1: a Ralstonia eutropha H16 megaplasmid encoding key enzymes of H(2)-based lithoautotrophy and anaerobiosis." Schwartz E., Henne A., Cramm R., Eitinger T., Friedrich B., Gottschalk G. J. Mol. Biol. 332:369-383(2003) [PubMed: 12948488] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
[3]	"Yeast flavohemoglobin is an ancient protein related to globins and a reductase family." Zhu H., Riggs A.F. Proc. Natl. Acad. Sci. U.S.A. 89:5015-5019(1992) [PubMed: 1594608] [Abstract] Cited for: PARTIAL PROTEIN SEQUENCE.
[4]	"An oxygen-binding flavohemoprotein from Alcaligenes eutrophus." Probst I., Wolf G., Schlegel H.G. Biochim. Biophys. Acta 576:471-478(1979) [PubMed: 218634] [Abstract] Cited for: COFACTOR, INDUCTION, SPECTRAL PROPERTIES.
[5]	"Nitric-oxide dioxygenase activity and function of flavohemoglobins. Sensitivity to nitric oxide and carbon monoxide inhibition." Gardner P.R., Gardner A.M., Martin L.A., Dou Y., Li T., Olson J.S., Zhu H., Riggs A.F. J. Biol. Chem. 275:31581-31587(2000) [PubMed: 10922365] [Abstract] Cited for: ENZYME ACTIVITY.
[6]	"Crystal structure of the flavohemoglobin from Alcaligenes eutrophus at 1.75-A resolution." Ermler U., Siddiqui R.A., Cramm R., Friedrich B. EMBO J. 14:6067-6077(1995) [PubMed: 8557026] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS).
[7]	"Phospholipid bound to the flavohemoprotein from Alcaligenes eutrophus." Ollesch G., Kaunzinger A., Juchelka D., Schubert-Zsilavecz M., Ermler U. Eur. J. Biochem. 262:396-405(1999) [PubMed: 10336624] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF NATIVE PROTEIN AND MUTANT V98F, PHOSPHOLIPID BINDING, MUTAGENESIS OF ALA-60 AND VAL-98.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

X74334 Genomic DNA. Translation: CAA52381.1.
AY305378 Genomic DNA. Translation: AAP85952.1.

PIR

A53396.

RefSeq

NP_942838.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1CQX	X-ray	1.75	A/B	1-403	[»]

ModBase

Search...

Genome annotation databases

GeneID

2656642.

GenomeReviews

Gene locus PHG200 in contig AY305378_GR.

KEGG

reh:PHG200.

Organism-specific databases

CMR

Search...

Phylogenomic databases

eggNOG

COG1018.

HOGENOM

HBG623097.

OMA

YSISCAP.

PhylomeDB

P39662.

Family and domain databases

HAMAP

MF_01252. Hmp.
[Tree]

InterPro

IPR017927. Fd_Rdtase_FAD-bd.
IPR012292. Globin.
IPR009050. Globin-like.
IPR000971. Globin_subset.
IPR008333. OxRdtase_FAD-bd_dom.
IPR001433. OxRdtase_FAD/NAD_bd.
IPR001221. Phe_hydroxylase.
IPR017938. Riboflavin_synthase-like_b-brl.
[Graphical view]

Gene3D

G3DSA:1.10.490.10. Globin_related. 1 hit.

Pfam

PF00970. FAD_binding_6. 1 hit.
PF00042. Globin. 1 hit.
PF00175. NAD_binding_1. 1 hit.
[Graphical view]

PRINTS

PR00410. PHEHYDRXLASE.

PROSITE

PS51384. FAD_FR. 1 hit.
PS01033. GLOBIN. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

HMP_RALEH

Accession

Primary (citable) accession number: P39662
Secondary accession number(s): Q7WXD4

Entry history

Integrated into UniProtKB/Swiss-Prot:	February 1, 1995
Last sequence update:	January 23, 2007
Last modified:	January 19, 2010
This is version 94 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)