PRND_PSEFL
ID PRND_PSEFL Reviewed; 363 AA.
AC P95483;
DT 29-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-1997, sequence version 1.
DT 03-AUG-2022, entry version 79.
DE RecName: Full=Aminopyrrolnitrin oxygenase PrnD;
DE EC=1.14.13.-;
DE AltName: Full=Arylamine oxygenase;
GN Name=prnD;
OS Pseudomonas fluorescens.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=294;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, DISRUPTION PHENOTYPE, AND
RP NOMENCLATURE.
RC STRAIN=Bl915;
RX PubMed=9172332; DOI=10.1128/aem.63.6.2147-2154.1997;
RA Hammer P.E., Hill D.S., Lam S.T., Van Pee K.H., Ligon J.M.;
RT "Four genes from Pseudomonas fluorescens that encode the biosynthesis of
RT pyrrolnitrin.";
RL Appl. Environ. Microbiol. 63:2147-2154(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=Bl915;
RX PubMed=9537395; DOI=10.1128/jb.180.7.1939-1943.1998;
RA Kirner S., Hammer P.E., Hill D.S., Altmann A., Fischer I., Weislo L.J.,
RA Lanahan M., van Pee K.H., Ligon J.M.;
RT "Functions encoded by pyrrolnitrin biosynthetic genes from Pseudomonas
RT fluorescens.";
RL J. Bacteriol. 180:1939-1943(1998).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP CYS-69; CYS-88; ASP-183 AND ASP-323, COFACTOR, SUBSTRATE SPECIFICITY, AND
RP SUBUNIT.
RX PubMed=16150698; DOI=10.1074/jbc.m505334200;
RA Lee J., Simurdiak M., Zhao H.;
RT "Reconstitution and characterization of aminopyrrolnitrin oxygenase, a
RT Rieske N-oxygenase that catalyzes unusual arylamine oxidation.";
RL J. Biol. Chem. 280:36719-36727(2005).
RN [4]
RP FUNCTION, MUTAGENESIS OF TRP-209; ASP-269 AND PHE-312, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBSTRATE SPECIFICITY, AND REACTION MECHANISM.
RX PubMed=16342311; DOI=10.1002/anie.200502903;
RA Lee J., Zhao H.;
RT "Mechanistic studies on the conversion of arylamines into arylnitro
RT compounds by aminopyrrolnitrin oxygenase: identification of intermediates
RT and kinetic studies.";
RL Angew. Chem. Int. Ed. 45:622-625(2006).
CC -!- FUNCTION: Involved in the biosynthesis of the antifungal antibiotic
CC pyrrolnitrin (PRN). Catalyzes the oxidation of the amino group of
CC aminopyrrolnitrin (APRN) to a nitro group to form PRN. It has high
CC substrate specificity toward physiological substrate aminopyrrolnitrin,
CC p-aminobenzylamine (pABA), p-aminobenzyl alcohol, and p-aminophenyl
CC alanine. {ECO:0000269|PubMed:16150698, ECO:0000269|PubMed:16342311,
CC ECO:0000269|PubMed:9172332, ECO:0000269|PubMed:9537395}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=aminopyrrolnitrin + H(+) + NADPH + 2 O2 = 2 H2O + NADP(+) +
CC pyrrolnitrin; Xref=Rhea:RHEA:46136, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:32079,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:85786;
CC Evidence={ECO:0000269|PubMed:16150698};
CC -!- COFACTOR:
CC Name=[2Fe-2S] cluster; Xref=ChEBI:CHEBI:190135;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU00628,
CC ECO:0000269|PubMed:16150698};
CC Note=Binds 1 [2Fe-2S] cluster per subunit. {ECO:0000255|PROSITE-
CC ProRule:PRU00628, ECO:0000269|PubMed:16150698};
CC -!- COFACTOR:
CC Name=Fe cation; Xref=ChEBI:CHEBI:24875;
CC Evidence={ECO:0000269|PubMed:16150698};
CC Note=Binds 1 Fe cation per subunit. {ECO:0000269|PubMed:16150698};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000269|PubMed:16150698};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=35.5 uM for p-aminophenyl alanine (at pH 7.8 and 30 degrees
CC Celsius) {ECO:0000269|PubMed:16150698, ECO:0000269|PubMed:16342311};
CC KM=42.2 uM for p-aminobenzyl alcohol (at pH 7.8 and 30 degrees
CC Celsius) {ECO:0000269|PubMed:16150698, ECO:0000269|PubMed:16342311};
CC KM=191 uM for APRN (at pH 7.8 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:16150698, ECO:0000269|PubMed:16342311};
CC KM=379 uM for pABA (at pH 7.8 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:16150698, ECO:0000269|PubMed:16342311};
CC Note=kcat is 6.8 min(-1) for aminopyrrolnitrin, 6.5 min(-1) for p-
CC aminobenzyl amine, 1.8 min(-1) for p-aminobenzyl alcohol and 1.2
CC min(-1) for p-aminophenyl alanine.;
CC -!- PATHWAY: Antibiotic biosynthesis.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:16150698}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene lose the ability to
CC produce pyrrolnitrin and accumulate APRN. {ECO:0000269|PubMed:9172332,
CC ECO:0000269|PubMed:9537395}.
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; U74493; AAB97507.1; -; Genomic_DNA.
DR AlphaFoldDB; P95483; -.
DR SMR; P95483; -.
DR PRIDE; P95483; -.
DR KEGG; ag:AAB97507; -.
DR GO; GO:0051537; F:2 iron, 2 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0008203; P:cholesterol metabolic process; IEA:InterPro.
DR CDD; cd03537; Rieske_RO_Alpha_PrnD; 1.
DR Gene3D; 2.102.10.10; -; 1.
DR InterPro; IPR045605; KshA-like_C.
DR InterPro; IPR037338; PrnD_Rieske.
DR InterPro; IPR017941; Rieske_2Fe-2S.
DR InterPro; IPR036922; Rieske_2Fe-2S_sf.
DR Pfam; PF19298; KshA_C; 1.
DR Pfam; PF00355; Rieske; 1.
DR SUPFAM; SSF50022; SSF50022; 1.
DR PROSITE; PS51296; RIESKE; 1.
PE 1: Evidence at protein level;
KW 2Fe-2S; Antibiotic biosynthesis; Flavoprotein; FMN; Iron; Iron-sulfur;
KW Metal-binding; NADP; Oxidoreductase.
FT CHAIN 1..363
FT /note="Aminopyrrolnitrin oxygenase PrnD"
FT /id="PRO_0000422333"
FT DOMAIN 29..137
FT /note="Rieske"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 69
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000305"
FT BINDING 71
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT BINDING 88
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000305"
FT BINDING 91
FT /ligand="[2Fe-2S] cluster"
FT /ligand_id="ChEBI:CHEBI:190135"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00628"
FT SITE 183
FT /note="Important for catalysis"
FT SITE 323
FT /note="Important for catalysis"
FT MUTAGEN 69
FT /note="C->A: Loss of oxygenase activity."
FT /evidence="ECO:0000269|PubMed:16150698"
FT MUTAGEN 88
FT /note="C->A: Loss of oxygenase activity."
FT /evidence="ECO:0000269|PubMed:16150698"
FT MUTAGEN 183
FT /note="D->A: Significant decrease of oxygenase activity."
FT /evidence="ECO:0000269|PubMed:16150698"
FT MUTAGEN 209
FT /note="W->A: Unable to convert either the alternative
FT substrate pABA or the interdiate 4-hydroxylaminobenzylamine
FT (pHABA)."
FT /evidence="ECO:0000269|PubMed:16342311"
FT MUTAGEN 269
FT /note="D->A: Shows much higher activity toward the
FT intermediate 4-hydroxylaminobenzylamine (pHABA) than pABA."
FT /evidence="ECO:0000269|PubMed:16342311"
FT MUTAGEN 312
FT /note="F->V: Unable to convert either the alternative
FT substrate pABA or the interdiate 4-hydroxylaminobenzylamine
FT (pHABA)."
FT /evidence="ECO:0000269|PubMed:16342311"
FT MUTAGEN 323
FT /note="D->A: Significant decrease of oxygenase."
FT /evidence="ECO:0000269|PubMed:16150698"
SQ SEQUENCE 363 AA; 40694 MW; 1EB056F18F85A560 CRC64;
MNDIQLDQAS VKKRPSGAYD ATTRLAASWY VAMRSNELKD KPTELTLFGR PCVAWRGATG
RAVVMDRHCS HLGANLADGR IKDGCIQCPF HHWRYDEQGQ CVHIPGHNQA VRQLEPVPRG
ARQPTLVTAE RYGYVWVWYG SPLPLHPLPE ISAADVDNGD FMHLHFAFET TTAVLRIVEN
FYDAQHATPV HALPISAFEL KLFDDWRQWP EVESLALAGA WFGAGIDFTV DRYFGPLGML
SRALGLNMSQ MNLHFDGYPG GCVMTVALDG DVKYKLLQCV TPVSEGKNVM HMLISIKKVG
GILRRATDFV LFGLQTRQAA GYDVKIWNGM KPDGGGAYSK YDKLVLKYRA FYRGWVDRVA
SER
