PRNA_PSEFL
ID PRNA_PSEFL Reviewed; 538 AA.
AC P95480;
DT 29-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-1997, sequence version 1.
DT 03-AUG-2022, entry version 76.
DE RecName: Full=Flavin-dependent tryptophan halogenase PrnA;
DE EC=1.14.19.9;
GN Name=prnA;
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, CATALYTIC ACTIVITY, AND
RP 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.
RX PubMed=10941070;
RX DOI=10.1002/1521-3773(20000703)39:13<2300::aid-anie2300>3.0.co;2-i;
RA Keller S., Wage T., Hohaus K., Holzer M., Eichhorn E., van Pee K.H.;
RT "Purification and partial characterization of tryptophan 7-halogenase
RT (PrnA) from Pseudomonas fluorescens.";
RL Angew. Chem. Int. Ed. 39:2300-2302(2000).
RN [4]
RP CRYSTALLIZATION.
RX PubMed=15272170; DOI=10.1107/s0907444904012521;
RA Dong C., Kotzsch A., Dorward M., van Pee K.H., Naismith J.H.;
RT "Crystallization and X-ray diffraction of a halogenating enzyme, tryptophan
RT 7-halogenase, from Pseudomonas fluorescens.";
RL Acta Crystallogr. D 60:1438-1440(2004).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEX WITH FAD, CHLORIDE AND
RP TRYPTOPHAN, ACTIVE SITE, MUTAGENESIS OF LYS-79 AND GLU-346, AND SUBUNIT.
RX PubMed=16195462; DOI=10.1126/science.1116510;
RA Dong C., Flecks S., Unversucht S., Haupt C., van Pee K.H., Naismith J.H.;
RT "Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for
RT regioselective chlorination.";
RL Science 309:2216-2219(2005).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF MUTANT ASP-346 IN COMPLEX WITH
RP FAD AND TRYPTOPHAN, MUTAGENESIS OF TRP-272; TRP-274; GLU-346 AND SER-347,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND REACTION MECHANISM.
RX PubMed=18979475; DOI=10.1002/anie.200802466;
RA Flecks S., Patallo E.P., Zhu X., Ernyei A.J., Seifert G., Schneider A.,
RA Dong C., Naismith J.H., van Pee K.H.;
RT "New insights into the mechanism of enzymatic chlorination of tryptophan.";
RL Angew. Chem. Int. Ed. 47:9533-9536(2008).
CC -!- FUNCTION: Involved in the biosynthesis of the antifungal antibiotic
CC pyrrolnitrin. Catalyzes the chlorination of tryptophan (Trp) at C7
CC position to yield 7-chloro-L-tryptophan (7-CLT). The reaction between
CC FADH2, Cl-, and O2 generates the powerful oxidant HOCl, which is
CC presumed to carry out the chlorination reaction. The reaction of HOCl
CC with the active site Lys-79 generates a lysine chloramine, which plays
CC a key role in directing regiospecific chlorination of substrate in this
CC important class of biosynthetic enzymes. It is also able to use bromide
CC ions to generate monobrominated Trp. {ECO:0000269|PubMed:10941070,
CC ECO:0000269|PubMed:9172332, ECO:0000269|PubMed:9537395}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=chloride + FADH2 + L-tryptophan + O2 = 7-chloro-L-tryptophan +
CC FAD + 2 H2O; Xref=Rhea:RHEA:26494, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17996, ChEBI:CHEBI:57692,
CC ChEBI:CHEBI:57912, ChEBI:CHEBI:58307, ChEBI:CHEBI:58713;
CC EC=1.14.19.9; Evidence={ECO:0000269|PubMed:9537395};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=17.1 uM for tryptophan (with FAD at pH 7.2 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:18979475};
CC Vmax=43 pmol/min/mg enzyme {ECO:0000269|PubMed:18979475};
CC -!- PATHWAY: Antibiotic biosynthesis.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:16195462,
CC ECO:0000269|PubMed:18979475}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene lose the ability to
CC produce pyrrolnitrin. {ECO:0000269|PubMed:9172332,
CC ECO:0000269|PubMed:9537395}.
CC -!- SIMILARITY: Belongs to the flavin-dependent halogenase family.
CC Bacterial tryptophan halogenase subfamily. {ECO:0000305}.
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DR EMBL; U74493; AAB97504.1; -; Genomic_DNA.
DR PDB; 2APG; X-ray; 1.90 A; A=1-538.
DR PDB; 2AQJ; X-ray; 1.80 A; A=1-538.
DR PDB; 2AR8; X-ray; 2.20 A; A=1-538.
DR PDB; 2ARD; X-ray; 2.60 A; A=1-538.
DR PDB; 2JKC; X-ray; 2.30 A; A=1-538.
DR PDB; 4Z43; X-ray; 2.29 A; A=1-538.
DR PDB; 4Z44; X-ray; 2.20 A; A=1-538.
DR PDBsum; 2APG; -.
DR PDBsum; 2AQJ; -.
DR PDBsum; 2AR8; -.
DR PDBsum; 2ARD; -.
DR PDBsum; 2JKC; -.
DR PDBsum; 4Z43; -.
DR PDBsum; 4Z44; -.
DR AlphaFoldDB; P95480; -.
DR SMR; P95480; -.
DR PRIDE; P95480; -.
DR KEGG; ag:AAB97504; -.
DR BioCyc; MetaCyc:MON-16710; -.
DR BRENDA; 1.14.19.9; 5121.
DR SABIO-RK; P95480; -.
DR EvolutionaryTrace; P95480; -.
DR GO; GO:0004497; F:monooxygenase activity; IEA:InterPro.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR006905; Flavin_halogenase.
DR InterPro; IPR033856; Trp_halogen.
DR Pfam; PF04820; Trp_halogenase; 1.
DR PIRSF; PIRSF011396; Trp_halogenase; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; FAD; Flavoprotein;
KW Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..538
FT /note="Flavin-dependent tryptophan halogenase PrnA"
FT /id="PRO_0000422330"
FT ACT_SITE 79
FT /evidence="ECO:0000269|PubMed:16195462"
FT BINDING 13..16
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT BINDING 39..50
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT BINDING 79
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 187
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT BINDING 337
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT BINDING 346
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 348..349
FT /ligand="chloride"
FT /ligand_id="ChEBI:CHEBI:17996"
FT BINDING 350
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT BINDING 443..444
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 450
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT BINDING 454
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT SITE 79
FT /note="Role in guiding and activating HOCl"
FT SITE 346
FT /note="Role in stabilization and deprotonation of the
FT intermediate"
FT MUTAGEN 79
FT /note="K->A: Loss of halogenase activity."
FT /evidence="ECO:0000269|PubMed:16195462"
FT MUTAGEN 272
FT /note="W->A: No change in halogenase activity."
FT /evidence="ECO:0000269|PubMed:18979475"
FT MUTAGEN 272
FT /note="W->F: No change in halogenase activity."
FT /evidence="ECO:0000269|PubMed:18979475"
FT MUTAGEN 274
FT /note="W->A: No change in halogenase activity."
FT /evidence="ECO:0000269|PubMed:18979475"
FT MUTAGEN 274
FT /note="W->F: No change in halogenase activity."
FT /evidence="ECO:0000269|PubMed:18979475"
FT MUTAGEN 346
FT /note="E->D: Loss of halogenase activity."
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT MUTAGEN 346
FT /note="E->Q: The catalytic efficiency decreases by about
FT two orders of magnitude, however the binding affinity is
FT unchanged."
FT /evidence="ECO:0000269|PubMed:16195462,
FT ECO:0000269|PubMed:18979475"
FT MUTAGEN 347
FT /note="S->A: Does not completely abolish halogenase
FT activity."
FT /evidence="ECO:0000269|PubMed:18979475"
FT STRAND 7..11
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 15..27
FT /evidence="ECO:0007829|PDB:2AQJ"
FT TURN 28..30
FT /evidence="ECO:0007829|PDB:2JKC"
FT STRAND 33..38
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 40..42
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 54..58
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 60..63
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 67..70
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 71..74
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 77..79
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 81..86
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 88..90
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 98..104
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 108..113
FT /evidence="ECO:0007829|PDB:4Z44"
FT HELIX 114..123
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 130..134
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 138..142
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 159..162
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 164..177
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 181..184
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 187..192
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 194..196
FT /evidence="ECO:0007829|PDB:2ARD"
FT STRAND 198..203
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 208..210
FT /evidence="ECO:0007829|PDB:4Z44"
FT STRAND 212..216
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 219..221
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 223..228
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 233..235
FT /evidence="ECO:0007829|PDB:2AQJ"
FT TURN 237..239
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 244..251
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 254..257
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 261..267
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 269..278
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 281..288
FT /evidence="ECO:0007829|PDB:2AQJ"
FT TURN 290..292
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 295..306
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 315..318
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 322..325
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 327..329
FT /evidence="ECO:0007829|PDB:2AQJ"
FT STRAND 332..334
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 336..338
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 344..346
FT /evidence="ECO:0007829|PDB:2APG"
FT HELIX 349..362
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 371..397
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 405..412
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 418..429
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 440..445
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 447..452
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 457..467
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 476..480
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 482..505
FT /evidence="ECO:0007829|PDB:2AQJ"
FT HELIX 509..516
FT /evidence="ECO:0007829|PDB:2AQJ"
SQ SEQUENCE 538 AA; 61075 MW; 03AAAC041A00CE7B CRC64;
MNKPIKNIVI VGGGTAGWMA ASYLVRALQQ QANITLIESA AIPRIGVGEA TIPSLQKVFF
DFLGIPEREW MPQVNGAFKA AIKFVNWRKS PDPSRDDHFY HLFGNVPNCD GVPLTHYWLR
KREQGFQQPM EYACYPQPGA LDGKLAPCLS DGTRQMSHAW HFDAHLVADF LKRWAVERGV
NRVVDEVVDV RLNNRGYISN LLTKEGRTLE ADLFIDCSGM RGLLINQALK EPFIDMSDYL
LCDSAVASAV PNDDARDGVE PYTSSIAMNS GWTWKIPMLG RFGSGYVFSS HFTSRDQATA
DFLKLWGLSD NQPLNQIKFR VGRNKRAWVN NCVSIGLSSC FLEPLESTGI YFIYAALYQL
VKHFPDTSFD PRLSDAFNAE IVHMFDDCRD FVQAHYFTTS RDDTPFWLAN RHDLRLSDAI
KEKVQRYKAG LPLTTTSFDD STYYETFDYE FKNFWLNGNY YCIFAGLGML PDRSLPLLQH
RPESIEKAEA MFASIRREAE RLRTSLPTNY DYLRSLRDGD AGLSRGQRGP KLAAQESL
