PRHA_PENBI
ID PRHA_PENBI Reviewed; 301 AA.
AC A0A1E1FFL0;
DT 26-FEB-2020, integrated into UniProtKB/Swiss-Prot.
DT 18-JAN-2017, sequence version 1.
DT 03-AUG-2022, entry version 18.
DE RecName: Full=Multifunctional dioxygenase prhA {ECO:0000303|PubMed:27602587};
DE EC=1.14.11.- {ECO:0000269|PubMed:27602587, ECO:0000269|PubMed:29317628};
DE AltName: Full=Paraherquonin biosynthesis cluster protein A {ECO:0000303|PubMed:27602587};
GN Name=prhA {ECO:0000303|PubMed:27602587};
OS Penicillium brasilianum.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=104259;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, AND
RP PATHWAY.
RC STRAIN=ATCC 22354 / NBRC 6234 / CBS 338.59 / FRR 3454 / IMI 68220;
RX PubMed=27602587; DOI=10.1021/jacs.6b08424;
RA Matsuda Y., Iwabuchi T., Fujimoto T., Awakawa T., Nakashima Y., Mori T.,
RA Zhang H., Hayashi F., Abe I.;
RT "Discovery of key dioxygenases that diverged the paraherquonin and
RT acetoxydehydroaustin pathways in Penicillium brasilianum.";
RL J. Am. Chem. Soc. 138:12671-12677(2016).
RN [2]
RP FUNCTION.
RX PubMed=28759016; DOI=10.1038/nchembio.2443;
RA Mori T., Iwabuchi T., Hoshino S., Wang H., Matsuda Y., Abe I.;
RT "Molecular basis for the unusual ring reconstruction in fungal
RT meroterpenoid biogenesis.";
RL Nat. Chem. Biol. 13:1066-1073(2017).
RN [3] {ECO:0007744|PDB:5YBM, ECO:0007744|PDB:5YBN, ECO:0007744|PDB:5YBO, ECO:0007744|PDB:5YBP, ECO:0007744|PDB:5YBQ, ECO:0007744|PDB:5YBR, ECO:0007744|PDB:5YBS, ECO:0007744|PDB:5YBT}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 6-294, SUBUNIT, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF VAL-150
RP AND ALA-232, AND PATHWAY.
RX PubMed=29317628; DOI=10.1038/s41467-017-02371-w;
RA Nakashima Y., Mori T., Nakamura H., Awakawa T., Hoshino S., Senda M.,
RA Senda T., Abe I.;
RT "Structure function and engineering of multifunctional non-heme iron
RT dependent oxygenases in fungal meroterpenoid biosynthesis.";
RL Nat. Commun. 9:104-104(2018).
CC -!- FUNCTION: Multifunctional dioxygenase; part of the gene cluster that
CC mediates the biosynthesis of paraherquonin, a meroterpenoid with a
CC unique, highly congested hexacyclic molecular architecture
CC (PubMed:27602587). The first step of the pathway is the synthesis of
CC 3,5-dimethylorsellinic acid (DMOA) by the polyketide synthase prhL (By
CC similarity). Synthesis of DMOA is followed by farnesylation by the
CC prenyltransferase prhE, methylesterification by the methyl-transferase
CC prhM, epoxidation of the prenyl chain by the flavin-dependent
CC monooxygenase prhF, and cyclization of the farnesyl moiety by the
CC terpene cyclase prhH, to yield the tetracyclic intermediate,
CC protoaustinoid A (By similarity). The short chain dehydrogenase prhI
CC then oxidizes the C-3 alcohol group of the terpene cyclase product to
CC transform protoaustinoid A into protoaustinoid B (PubMed:27602587). The
CC FAD-binding monooxygenase prhJ catalyzes the oxidation of
CC protoaustinoid B into preaustinoid A which is further oxidized into
CC preaustinoid A1 by FAD-binding monooxygenase phrK (PubMed:27602587).
CC Finally, prhA leads to berkeleydione via the berkeleyone B intermediate
CC (PubMed:27602587, PubMed:29317628). PrhA is a multifunctional
CC dioxygenase that first desaturates at C5-C6 to form berkeleyone B,
CC followed by rearrangement of the A/B-ring to form the cycloheptadiene
CC moiety in berkeleydione (PubMed:27602587, PubMed:29317628).
CC Berkeleydione serves as the key intermediate for the biosynthesis of
CC paraherquonin as well as many other meroterpenoids (Probable). The
CC cytochrome P450 monooxygenases prhB, prhD, and prhN, as well as the
CC isomerase prhC, are probably involved in the late stage of
CC paraherquonin biosynthesis, after the production of berkeleydione
CC (Probable). Especially prhC might be a multifunctional enzyme that
CC catalyzes the D-ring expansion via intramolecular methoxy
CC rearrangement, as well as the hydrolysis of the expanded D-ring
CC (Probable). {ECO:0000250|UniProtKB:Q5ATJ7, ECO:0000269|PubMed:27602587,
CC ECO:0000269|PubMed:29317628, ECO:0000305|PubMed:27602587,
CC ECO:0000305|PubMed:28759016, ECO:0000305|PubMed:29317628}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + O2 + preaustinoid A1 = berkeleyone B + CO2 +
CC H2O + succinate; Xref=Rhea:RHEA:65184, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16526, ChEBI:CHEBI:16810,
CC ChEBI:CHEBI:30031, ChEBI:CHEBI:69025, ChEBI:CHEBI:69026;
CC Evidence={ECO:0000269|PubMed:27602587, ECO:0000269|PubMed:29317628};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:65185;
CC Evidence={ECO:0000269|PubMed:27602587, ECO:0000269|PubMed:29317628};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + berkeleyone B + O2 = berkeleydione + CO2 +
CC H2O + succinate; Xref=Rhea:RHEA:65188, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16526, ChEBI:CHEBI:16810,
CC ChEBI:CHEBI:30031, ChEBI:CHEBI:69021, ChEBI:CHEBI:69025;
CC Evidence={ECO:0000269|PubMed:27602587};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:65189;
CC Evidence={ECO:0000269|PubMed:27602587};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 2-oxoglutarate + 2 O2 + preaustinoid A = berkeleytrione + 2
CC CO2 + H2O + 2 succinate; Xref=Rhea:RHEA:65192, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16526, ChEBI:CHEBI:16810,
CC ChEBI:CHEBI:30031, ChEBI:CHEBI:69022, ChEBI:CHEBI:69023;
CC Evidence={ECO:0000269|PubMed:27602587};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:65193;
CC Evidence={ECO:0000269|PubMed:27602587};
CC -!- COFACTOR:
CC Name=Fe cation; Xref=ChEBI:CHEBI:24875;
CC Evidence={ECO:0000250|UniProtKB:Q4WAW9};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=48.3 uM for preaustinoid A1 {ECO:0000269|PubMed:29317628};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000269|PubMed:27602587, ECO:0000269|PubMed:29317628}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:29317628}.
CC -!- DOMAIN: Residues at positions 150 and 232 are important for type of
CC reaction catalyzed, using identical substrate (PubMed:29317628). Both
CC ausE and prhA accept preaustinoid A1 as a substrate to form divergent
CC products through dynamic skeletal rearrangement. AusE (containing Leu-
CC 150 and Ser-232) first desaturates at C1-C2 to form preaustinoid
CC A2,followed by rearrangement leading to the formation of the spiro-
CC lactone in preaustinoid A3 (PubMed:29317628). In contrast, prhA
CC (containing Val-150 and Ala-232) first desaturates at C5-C6 to form
CC berkeleyone B, followed by rearrangement of the A/B-ring to form the
CC cycloheptadiene moiety in berkeleydione (PubMed:29317628).
CC {ECO:0000269|PubMed:29317628}.
CC -!- SIMILARITY: Belongs to the PhyH 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; LC127182; BAV69302.1; -; Genomic_DNA.
DR PDB; 5YBM; X-ray; 2.12 A; A/B=6-294.
DR PDB; 5YBN; X-ray; 2.10 A; A/B=6-294.
DR PDB; 5YBO; X-ray; 2.20 A; A/B=6-294.
DR PDB; 5YBP; X-ray; 2.31 A; A/B=6-294.
DR PDB; 5YBQ; X-ray; 2.25 A; A/B=6-294.
DR PDB; 5YBR; X-ray; 2.26 A; A/B=6-294.
DR PDB; 5YBS; X-ray; 2.30 A; A/B=6-294.
DR PDB; 5YBT; X-ray; 2.35 A; A/B=6-294.
DR PDBsum; 5YBM; -.
DR PDBsum; 5YBN; -.
DR PDBsum; 5YBO; -.
DR PDBsum; 5YBP; -.
DR PDBsum; 5YBQ; -.
DR PDBsum; 5YBR; -.
DR PDBsum; 5YBS; -.
DR PDBsum; 5YBT; -.
DR AlphaFoldDB; A0A1E1FFL0; -.
DR SMR; A0A1E1FFL0; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0051213; F:dioxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniPathway.
DR InterPro; IPR008775; Phytyl_CoA_dOase.
DR Pfam; PF05721; PhyH; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Dioxygenase; Iron; Metal-binding; Oxidoreductase.
FT CHAIN 1..301
FT /note="Multifunctional dioxygenase prhA"
FT /id="PRO_0000449162"
FT BINDING 130
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:29317628"
FT BINDING 132
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:29317628"
FT BINDING 214
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:29317628"
FT SITE 150
FT /note="Important for reaction specificity"
FT /evidence="ECO:0000269|PubMed:29317628"
FT SITE 232
FT /note="Important for reaction specificity"
FT /evidence="ECO:0000269|PubMed:29317628"
FT MUTAGEN 150
FT /note="V->L: Completely loses its original activity, and
FT instead catalyzes the AusE-type 'spiro-lactone formation'
FT to yield preaustinoid A3 as a single product; when
FT associated with S-232."
FT /evidence="ECO:0000269|PubMed:29317628"
FT MUTAGEN 232
FT /note="A->S: Completely loses its original activity, and
FT instead catalyzes the AusE-type 'spiro-lactone formation'
FT to yield preaustinoid A3 as a single product; when
FT associated with L-150."
FT /evidence="ECO:0000269|PubMed:29317628"
FT STRAND 11..13
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 18..28
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 29..34
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 39..55
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 69..73
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 76..79
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 81..85
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 87..89
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 91..101
FT /evidence="ECO:0007829|PDB:5YBN"
FT TURN 104..106
FT /evidence="ECO:0007829|PDB:5YBM"
FT STRAND 110..120
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 132..135
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 139..145
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 150..158
FT /evidence="ECO:0007829|PDB:5YBN"
FT TURN 162..165
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 168..170
FT /evidence="ECO:0007829|PDB:5YBO"
FT HELIX 173..175
FT /evidence="ECO:0007829|PDB:5YBN"
FT TURN 184..186
FT /evidence="ECO:0007829|PDB:5YBN"
FT TURN 188..191
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 205..209
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 212..215
FT /evidence="ECO:0007829|PDB:5YBO"
FT STRAND 226..235
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 244..247
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 250..254
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 258..263
FT /evidence="ECO:0007829|PDB:5YBN"
FT STRAND 269..272
FT /evidence="ECO:0007829|PDB:5YBM"
FT STRAND 279..283
FT /evidence="ECO:0007829|PDB:5YBN"
FT HELIX 288..292
FT /evidence="ECO:0007829|PDB:5YBN"
SQ SEQUENCE 301 AA; 33731 MW; 0C37B8CFA7288904 CRC64;
MAPMIPPRLQ RFPATASADE IFAAFQEDGC VVIEGFISPE QVARFSQEVD PAMEKIPVEV
TNNGNSNDRT KRFSKCVIAS PTFRNEIIES DLMHELCDRV FSKPGEGMGY HFNDNMVIEV
QPGAPAQRLH RDQELYPWWN SMGPAGPECV INFFCAVTPF TEENGATRLV PGSHLWPEFT
QINERDCPQF GKIETVPAIM QPGDCYLMSG KVIHGAGHNA TTTDRRRALA LAIIRRELRP
MQAFSLSVPM KLAREMSERS QTMFGFRSSV QHCDVDMVHF WGNDGKDIAH HLGLEAPSVH
V
