NP4_RHOPR
ID NP4_RHOPR Reviewed; 205 AA.
AC Q94734;
DT 15-JUL-1998, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 03-AUG-2022, entry version 110.
DE RecName: Full=Nitrophorin-4 {ECO:0000303|Ref.1};
DE Short=NP4 {ECO:0000303|Ref.1};
DE EC=1.7.6.1 {ECO:0000269|PubMed:20524697};
DE AltName: Full=Nitrite dismutase {ECO:0000305};
DE Flags: Precursor;
OS Rhodnius prolixus (Triatomid bug).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Paraneoptera; Hemiptera; Heteroptera; Panheteroptera;
OC Cimicomorpha; Reduviidae; Triatominae; Rhodnius.
OX NCBI_TaxID=13249;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RC TISSUE=Salivary gland;
RA Champagne D.E., Ribeiro J.M.C.;
RL Submitted (SEP-1996) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION.
RX PubMed=15598503; DOI=10.1016/j.jinorgbio.2004.10.009;
RA Walker F.A.;
RT "Nitric oxide interaction with insect nitrophorins and thoughts on the
RT electron configuration of the {FeNO}6 complex.";
RL J. Inorg. Biochem. 99:216-236(2005).
RN [3]
RP DISULFIDE BOND.
RX PubMed=21955842; DOI=10.1016/j.jinorgbio.2011.07.009;
RA Knipp M., Taing J.J., He C.;
RT "Reduction of the lipocalin type heme containing protein nitrophorin
RT -- sensitivity of the fold-stabilizing cysteine disulfides toward routine
RT heme-iron reduction.";
RL J. Inorg. Biochem. 105:1405-1412(2011).
RN [4]
RP FUNCTION.
RX PubMed=23133364; DOI=10.1371/journal.pcbi.1002761;
RA Di Russo N.V., Estrin D.A., Marti M.A., Roitberg A.E.;
RT "pH-Dependent conformational changes in proteins and their effect on
RT experimental pK(a)s: the case of Nitrophorin 4.";
RL PLoS Comput. Biol. 8:E1002761-E1002761(2012).
RN [5]
RP FUNCTION.
RX PubMed=23885811; DOI=10.1021/jp407052a;
RA Cheng M., Brookes J.F., Montfort W.R., Khalil M.;
RT "pH-dependent picosecond structural dynamics in the distal pocket of
RT nitrophorin 4 investigated by 2D IR spectroscopy.";
RL J. Phys. Chem. B 117:15804-15811(2013).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME.
RX PubMed=9782054; DOI=10.1016/s0969-2126(98)00131-2;
RA Andersen J.F., Weichsel A., Balfour C.A., Champagne D.E., Montfort W.R.;
RT "The crystal structure of nitrophorin 4 at 1.5-A resolution: transport of
RT nitric oxide by a lipocalin-based heme protein.";
RL Structure 6:1315-1327(1998).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME AND
RP NITRIC OXIDE.
RX PubMed=10876239; DOI=10.1038/76769;
RA Weichsel A., Andersen J.F., Roberts S.A., Montfort W.R.;
RT "Nitric oxide binding to nitrophorin 4 induces complete distal pocket
RT burial.";
RL Nat. Struct. Biol. 7:551-554(2000).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.08 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME;
RP NITRIC OXIDE; AMMONIA; HISTAMINE AND IMIDAZOLE.
RX PubMed=11560480; DOI=10.1021/bi0109257;
RA Roberts S.A., Weichsel A., Qiu Y., Shelnutt J.A., Walker F.A.,
RA Montfort W.R.;
RT "Ligand-induced heme ruffling and bent NO geometry in ultra-high-resolution
RT structures of nitrophorin 4.";
RL Biochemistry 40:11327-11337(2001).
RN [9]
RP FUNCTION, X-RAY CRYSTALLOGRAPHY (1.00 ANGSTROMS) OF 22-205 IN COMPLEX WITH
RP HEME; NITRIC OXIDE; AMMONIA AND IMIDAZOLE, AND MUTAGENESIS OF ASP-51;
RP THR-142 AND 150-ASP-LEU-151.
RX PubMed=15157102; DOI=10.1021/bi049748a;
RA Maes E.M., Weichsel A., Andersen J.F., Shepley D., Montfort W.R.;
RT "Role of binding site loops in controlling nitric oxide release: structure
RT and kinetics of mutant forms of nitrophorin 4.";
RL Biochemistry 43:6679-6690(2004).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (0.85 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME AND
RP NITRIC OXIDE, AND DISULFIDE BOND.
RX PubMed=15504026; DOI=10.1021/bi0483155;
RA Kondrashov D.A., Roberts S.A., Weichsel A., Montfort W.R.;
RT "Protein functional cycle viewed at atomic resolution: conformational
RT change and mobility in nitrophorin 4 as a function of pH and NO binding.";
RL Biochemistry 43:13637-13647(2004).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (1.45 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME.
RX PubMed=15258143; DOI=10.1074/jbc.m406178200;
RA Nienhaus K., Maes E.M., Weichsel A., Montfort W.R., Nienhaus G.U.;
RT "Structural dynamics controls nitric oxide affinity in nitrophorin 4.";
RL J. Biol. Chem. 279:39401-39407(2004).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (1.25 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME AND
RP 4-IODOPYRAZOLE.
RX PubMed=14673714; DOI=10.1007/s00775-003-0505-0;
RA Berry R.E., Ding X.D., Shokhireva T.K.H., Weichsel A., Montfort W.R.,
RA Walker F.A.;
RT "Axial ligand complexes of the Rhodnius nitrophorins: reduction potentials,
RT binding constants, EPR spectra, and structures of the 4-iodopyrazole and
RT imidazole complexes of NP4.";
RL J. Biol. Inorg. Chem. 9:135-144(2004).
RN [13]
RP X-RAY CRYSTALLOGRAPHY (0.89 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME;
RP NITRIC OXYDE AND CARBON MONOXIDE.
RX PubMed=16171383; DOI=10.1021/bi0506573;
RA Maes E.M., Roberts S.A., Weichsel A., Montfort W.R.;
RT "Ultrahigh resolution structures of nitrophorin 4: heme distortion in
RT ferrous CO and NO complexes.";
RL Biochemistry 44:12690-12699(2005).
RN [14]
RP X-RAY CRYSTALLOGRAPHY (1.11 ANGSTROMS) OF 22-205.
RX PubMed=17660249; DOI=10.1110/ps.072981907;
RA Amoia A.M., Montfort W.R.;
RT "Apo-nitrophorin 4 at atomic resolution.";
RL Protein Sci. 16:2076-2081(2007).
RN [15]
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME AND
RP NITRIC OXYDE, AND MUTAGENESIS OF ASP-51 AND GLU-76.
RX PubMed=19175316; DOI=10.1021/ja808105d;
RA Berry R.E., Shokhirev M.N., Ho A.Y., Yang F., Shokhireva T.K., Zhang H.,
RA Weichsel A., Montfort W.R., Walker F.A.;
RT "Effect of mutation of carboxyl side-chain amino acids near the heme on the
RT midpoint potentials and ligand binding constants of nitrophorin 2 and its
RT NO, histamine, and imidazole complexes.";
RL J. Am. Chem. Soc. 131:2313-2327(2009).
RN [16]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND X-RAY CRYSTALLOGRAPHY (1.40
RP ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME AND NITRITE.
RX PubMed=20524697; DOI=10.1021/bi100324z;
RA He C., Ogata H., Knipp M.;
RT "Formation of the complex of nitrite with the ferriheme b beta-barrel
RT proteins nitrophorin 4 and nitrophorin 7.";
RL Biochemistry 49:5841-5851(2010).
RN [17]
RP X-RAY CRYSTALLOGRAPHY (1.30 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME, AND
RP MUTAGENESIS OF LEU-151.
RX PubMed=22334402; DOI=10.1002/anie.201108691;
RA He C., Fuchs M.R., Ogata H., Knipp M.;
RT "Guanidine-ferroheme coordination in the mutant protein nitrophorin
RT 4(L130R).";
RL Angew. Chem. Int. Ed. Engl. 51:4470-4473(2012).
RN [18]
RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME AND
RP NITRITE, AND MUTAGENESIS OF ASP-51 AND LEU-151.
RX PubMed=22976968; DOI=10.1002/cbdv.201100401;
RA He C., Ogata H., Knipp M.;
RT "Insertion of an H-bonding residue into the distal pocket of the ferriheme
RT protein nitrophorin 4: effect on nitrite-iron coordination and nitrite
RT disproportionation.";
RL Chem. Biodivers. 9:1761-1775(2012).
RN [19]
RP X-RAY CRYSTALLOGRAPHY (1.30 ANGSTROMS) OF 22-205 IN COMPLEX WITH HEME;
RP L-CYSTEINE AND L-HOMOCYSTEINE.
RX PubMed=23474537; DOI=10.1016/j.jinorgbio.2013.01.012;
RA He C., Nishikawa K., Erdem O.F., Reijerse E., Ogata H., Lubitz W.,
RA Knipp M.;
RT "Complexes of ferriheme nitrophorin 4 with low-molecular weight thiol(ate)s
RT occurring in blood plasma.";
RL J. Inorg. Biochem. 122:38-48(2013).
CC -!- FUNCTION: Heme-based protein that delivers nitric oxide gas (NO) to the
CC victim while feeding, resulting in vasodilation and inhibition of
CC platelet aggregation. Also binds tightly to histamine, which is
CC released by the host to induce wound healing. NO release is pH
CC dependent and linked to loop dynamics. {ECO:0000269|PubMed:15157102,
CC ECO:0000269|PubMed:15598503, ECO:0000269|PubMed:20524697,
CC ECO:0000269|PubMed:23133364, ECO:0000269|PubMed:23885811}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 H(+) + 3 nitrite = H2O + nitrate + 2 nitric oxide;
CC Xref=Rhea:RHEA:31367, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16301, ChEBI:CHEBI:16480, ChEBI:CHEBI:17632; EC=1.7.6.1;
CC Evidence={ECO:0000269|PubMed:20524697};
CC -!- COFACTOR:
CC Name=heme b; Xref=ChEBI:CHEBI:60344;
CC Evidence={ECO:0000269|PubMed:20524697};
CC Note=Binds 1 heme b (iron(II)-protoporphyrin IX) group per subunit.
CC {ECO:0000269|PubMed:20524697};
CC -!- SUBCELLULAR LOCATION: Secreted.
CC -!- TISSUE SPECIFICITY: Expressed by the salivary glands.
CC -!- MISCELLANEOUS: Superposition of the structures of NP4 in the presence
CC of ammonia or nitrite shows a conformational change of the protein upon
CC NO(2)(-) binding. {ECO:0000269|PubMed:10876239,
CC ECO:0000269|PubMed:11560480, ECO:0000269|PubMed:15157102,
CC ECO:0000269|PubMed:15504026, ECO:0000269|PubMed:22976968}.
CC -!- SIMILARITY: Belongs to the calycin superfamily. Nitrophorin family.
CC {ECO:0000305}.
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DR EMBL; U70584; AAB09590.1; -; mRNA.
DR PIR; D56385; D56385.
DR PDB; 1D2U; X-ray; 1.15 A; A=22-205.
DR PDB; 1D3S; X-ray; 1.40 A; A=22-205.
DR PDB; 1EQD; X-ray; 1.60 A; A=22-205.
DR PDB; 1ERX; X-ray; 1.40 A; A=22-205.
DR PDB; 1IKE; X-ray; 1.50 A; A=22-205.
DR PDB; 1IKJ; X-ray; 1.27 A; A=22-205.
DR PDB; 1KOI; X-ray; 1.08 A; A=22-205.
DR PDB; 1ML7; X-ray; 1.25 A; A=22-205.
DR PDB; 1NP4; X-ray; 1.50 A; A=22-205.
DR PDB; 1SXU; X-ray; 1.40 A; A=22-205.
DR PDB; 1SXW; X-ray; 1.05 A; A=22-205.
DR PDB; 1SXX; X-ray; 1.01 A; A=22-205.
DR PDB; 1SXY; X-ray; 1.07 A; A=22-205.
DR PDB; 1SY0; X-ray; 1.15 A; A=22-205.
DR PDB; 1SY1; X-ray; 1.01 A; A=22-205.
DR PDB; 1SY2; X-ray; 1.00 A; A=22-205.
DR PDB; 1SY3; X-ray; 1.00 A; A=22-205.
DR PDB; 1U0X; X-ray; 1.45 A; A=22-205.
DR PDB; 1X8N; X-ray; 1.08 A; A=22-205.
DR PDB; 1X8O; X-ray; 1.01 A; A=22-205.
DR PDB; 1X8P; X-ray; 0.85 A; A=22-205.
DR PDB; 1X8Q; X-ray; 0.85 A; A=22-205.
DR PDB; 1YWA; X-ray; 0.89 A; A=22-205.
DR PDB; 1YWB; X-ray; 0.97 A; A=22-205.
DR PDB; 1YWC; X-ray; 1.00 A; A=22-205.
DR PDB; 1YWD; X-ray; 1.08 A; A=22-205.
DR PDB; 2AT0; X-ray; 1.00 A; X=22-205.
DR PDB; 2AT3; X-ray; 1.00 A; X=22-205.
DR PDB; 2AT5; X-ray; 1.22 A; X=22-205.
DR PDB; 2AT6; X-ray; 1.22 A; X=22-205.
DR PDB; 2AT8; X-ray; 1.00 A; X=22-205.
DR PDB; 2OFM; X-ray; 1.11 A; X=22-205.
DR PDB; 2OFR; X-ray; 1.00 A; X=22-205.
DR PDB; 3C76; X-ray; 1.07 A; X=22-205.
DR PDB; 3C77; X-ray; 1.08 A; X=22-205.
DR PDB; 3C78; X-ray; 0.98 A; X=22-205.
DR PDB; 3FLL; X-ray; 1.50 A; A=22-205.
DR PDB; 3MVF; X-ray; 1.40 A; A=22-205.
DR PDB; 3TGA; X-ray; 1.30 A; A=22-205.
DR PDB; 3TGB; X-ray; 1.35 A; A=22-205.
DR PDB; 3TGC; X-ray; 1.40 A; A=22-205.
DR PDB; 4GNW; X-ray; 1.15 A; A/B=22-205.
DR PDB; 4GRJ; X-ray; 1.15 A; A/B=22-205.
DR PDB; 4HPA; X-ray; 1.50 A; A=22-205.
DR PDB; 4HPB; X-ray; 1.60 A; A=22-205.
DR PDB; 4HPC; X-ray; 1.40 A; A=22-205.
DR PDB; 4HPD; X-ray; 1.30 A; A=22-205.
DR PDB; 5HWZ; X-ray; 1.45 A; A=22-205.
DR PDBsum; 1D2U; -.
DR PDBsum; 1D3S; -.
DR PDBsum; 1EQD; -.
DR PDBsum; 1ERX; -.
DR PDBsum; 1IKE; -.
DR PDBsum; 1IKJ; -.
DR PDBsum; 1KOI; -.
DR PDBsum; 1ML7; -.
DR PDBsum; 1NP4; -.
DR PDBsum; 1SXU; -.
DR PDBsum; 1SXW; -.
DR PDBsum; 1SXX; -.
DR PDBsum; 1SXY; -.
DR PDBsum; 1SY0; -.
DR PDBsum; 1SY1; -.
DR PDBsum; 1SY2; -.
DR PDBsum; 1SY3; -.
DR PDBsum; 1U0X; -.
DR PDBsum; 1X8N; -.
DR PDBsum; 1X8O; -.
DR PDBsum; 1X8P; -.
DR PDBsum; 1X8Q; -.
DR PDBsum; 1YWA; -.
DR PDBsum; 1YWB; -.
DR PDBsum; 1YWC; -.
DR PDBsum; 1YWD; -.
DR PDBsum; 2AT0; -.
DR PDBsum; 2AT3; -.
DR PDBsum; 2AT5; -.
DR PDBsum; 2AT6; -.
DR PDBsum; 2AT8; -.
DR PDBsum; 2OFM; -.
DR PDBsum; 2OFR; -.
DR PDBsum; 3C76; -.
DR PDBsum; 3C77; -.
DR PDBsum; 3C78; -.
DR PDBsum; 3FLL; -.
DR PDBsum; 3MVF; -.
DR PDBsum; 3TGA; -.
DR PDBsum; 3TGB; -.
DR PDBsum; 3TGC; -.
DR PDBsum; 4GNW; -.
DR PDBsum; 4GRJ; -.
DR PDBsum; 4HPA; -.
DR PDBsum; 4HPB; -.
DR PDBsum; 4HPC; -.
DR PDBsum; 4HPD; -.
DR PDBsum; 5HWZ; -.
DR AlphaFoldDB; Q94734; -.
DR SMR; Q94734; -.
DR STRING; 13249.RPRC000023-PA; -.
DR VEuPathDB; VectorBase:RPRC000023; -.
DR HOGENOM; CLU_117833_0_0_1; -.
DR BRENDA; 1.7.6.1; 5379.
DR EvolutionaryTrace; Q94734; -.
DR Proteomes; UP000015103; Unassembled WGS sequence.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0051381; F:histamine binding; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0070026; F:nitric oxide binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0042311; P:vasodilation; IEA:UniProtKB-KW.
DR Gene3D; 2.40.128.20; -; 1.
DR InterPro; IPR012674; Calycin.
DR InterPro; IPR023613; Nitrophorin.
DR InterPro; IPR002351; Nitrophorin_domain.
DR Pfam; PF02087; Nitrophorin; 1.
DR PRINTS; PR00788; NITROPHORIN.
DR SUPFAM; SSF50814; SSF50814; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Disulfide bond; Heme; Iron; Metal-binding; Oxidoreductase;
KW Reference proteome; Secreted; Signal; Vasoactive; Vasodilator.
FT SIGNAL 1..21
FT CHAIN 22..205
FT /note="Nitrophorin-4"
FT /id="PRO_0000021826"
FT BINDING 80
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="proximal binding residue"
FT /evidence="ECO:0000269|PubMed:10876239,
FT ECO:0000269|PubMed:11560480, ECO:0000269|PubMed:14673714,
FT ECO:0000269|PubMed:16171383, ECO:0000269|PubMed:20524697,
FT ECO:0000269|PubMed:22334402, ECO:0000269|PubMed:22976968,
FT ECO:0000269|PubMed:23474537, ECO:0000269|PubMed:9782054"
FT DISULFID 23..143
FT /evidence="ECO:0000269|PubMed:15504026,
FT ECO:0000269|PubMed:21955842"
FT DISULFID 62..192
FT /evidence="ECO:0000269|PubMed:15504026,
FT ECO:0000269|PubMed:21955842"
FT MUTAGEN 51
FT /note="D->A,N: Loss of NO-induced conformational change and
FT strongly reduced pH dependence for NO release.
FT Stabilization of the ferric-NO heme center and decreased
FT nitrite disproportionation."
FT /evidence="ECO:0000269|PubMed:15157102,
FT ECO:0000269|PubMed:19175316, ECO:0000269|PubMed:22976968"
FT MUTAGEN 76
FT /note="E->Q: Stabilization of the ferric-NO heme center."
FT /evidence="ECO:0000269|PubMed:19175316"
FT MUTAGEN 142
FT /note="T->V: No effect on NO-induced conformational change
FT or on pH dependence for NO release. Increased NO
FT association rates."
FT /evidence="ECO:0000269|PubMed:15157102"
FT MUTAGEN 150..151
FT /note="DL->AA: Loss of NO-induced conformational change and
FT loss of pH dependence for NO release."
FT /evidence="ECO:0000269|PubMed:15157102"
FT MUTAGEN 151
FT /note="L->R: Coordination of the heme iron center and
FT inhibition of nitrite disproportionation."
FT /evidence="ECO:0000269|PubMed:22334402,
FT ECO:0000269|PubMed:22976968"
FT HELIX 35..38
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 40..53
FT /evidence="ECO:0007829|PDB:1X8P"
FT HELIX 54..56
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 62..70
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 73..81
FT /evidence="ECO:0007829|PDB:1X8P"
FT TURN 83..85
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 88..99
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 102..110
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 116..118
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 124..133
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 135..146
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 153..161
FT /evidence="ECO:0007829|PDB:1X8P"
FT HELIX 168..176
FT /evidence="ECO:0007829|PDB:1X8P"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:1X8P"
FT STRAND 184..186
FT /evidence="ECO:0007829|PDB:1X8P"
FT HELIX 188..190
FT /evidence="ECO:0007829|PDB:1X8P"
FT HELIX 196..202
FT /evidence="ECO:0007829|PDB:1X8P"
SQ SEQUENCE 205 AA; 22406 MW; 9BAACA12001527F2 CRC64;
MKSYTSLLAV AILCLFGGVN GACTKNAIAQ TGFNKDKYFN GDVWYVTDYL DLEPDDVPKR
YCAALAAGTA SGKLKEALYH YDPKTQDTFY DVSELQVESL GKYTANFKKV DKNGNVKVAV
TAGNYYTFTV MYADDSSALI HTCLHKGNKD LGDLYAVLNR NKDAAAGDKV KSAVSAATLE
FSKFISTKEN NCAYDNDSLK SLLTK
