PLE3_RHOPP
ID PLE3_RHOPP Reviewed; 959 AA.
AC A0A6S6QR11;
DT 29-SEP-2021, integrated into UniProtKB/Swiss-Prot.
DT 02-DEC-2020, sequence version 1.
DT 03-AUG-2022, entry version 6.
DE RecName: Full=Bifunctional premutilin synthase {ECO:0000303|PubMed:28924980};
DE Short=PS {ECO:0000303|PubMed:28924980};
DE Includes:
DE RecName: Full=Class II diterpene cyclase {ECO:0000303|PubMed:28924980};
DE EC=5.5.1.- {ECO:0000303|PubMed:28924980};
DE Includes:
DE RecName: Full=Class I diterpene synthase {ECO:0000303|PubMed:28924980};
DE EC=4.2.3.- {ECO:0000303|PubMed:28924980};
GN Name=ple3 {ECO:0000303|PubMed:28924980};
OS Rhodocybe pseudopiperita (Clitopilus pseudopiperitus).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes;
OC Agaricomycetidae; Agaricales; Entolomataceae; Rhodocybe.
OX NCBI_TaxID=693819;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP MUTAGENESIS OF ASP-311 AND ASP-649, AND PATHWAY.
RC STRAIN=ATCC 20527;
RX PubMed=28924980; DOI=10.1002/cbic.201700434;
RA Yamane M., Minami A., Liu C., Ozaki T., Takeuchi I., Tsukagoshi T.,
RA Tokiwano T., Gomi K., Oikawa H.;
RT "Biosynthetic machinery of diterpene pleuromutilin isolated from
RT basidiomycete fungi.";
RL ChemBioChem 18:2317-2322(2017).
CC -!- FUNCTION: Bifunctional premutilin synthase; part of the gene cluster
CC that mediates the biosynthesis of pleuromutilin, a tricyclic diterpene
CC showing antibacterial properties (PubMed:28924980). The geranylgeranyl
CC diphosphate (GGPP) synthase ple4 catalyzes the first step in
CC pleuromutilin biosynthesis (PubMed:28924980). GGPP is then substrate of
CC the premutilin synthase (PS) ple3 to yield premutilin
CC (PubMed:28924980). Premutilin synthase is a bifunctional enzyme
CC composed of the fusion of a class II diterpene cyclase (DTC) and a
CC class I diterpene synthase (DTS), with the corresponding domains and
CC active sites containing characteristic aspartate-rich motifs (By
CC similarity). GGPP is first converted to mutildienyl-diphosphate (MPP)
CC at the class II DTC site (By similarity). MPP is subsequently further
CC cyclized at the class I DTS site, followed by a 1,5-hydride shift and
CC addition of water prior to terminating deprotonation, to yield
CC premutilin (By similarity). The cytochrome P450 monooxygenases ple5 and
CC ple6 hydroxylate premutilin at C-11 and C-3, respectively, producing
CC 11-hydroxypremutilin and 3-hydroxypremutilin (PubMed:28924980). The
CC combination of the actions of both ple5 and ple6 leads to the
CC production of 3,11-dihydroxypremutilin (PubMed:28924980). The short
CC chain dehydrogenase ple7 further converts 3,11-dihydroxypremutilin into
CC mutilin (PubMed:28924980). The acetyltransferase ple2 then acetylates
CC mutilin to produce 14-O-acetylmutilin (PubMed:28924980). Finally, the
CC cytochrome P450 monooxygenase ple1 catalyzes hydroxylation on the alpha
CC position of the acetyl side chain of 14-O-acetylmutilin to yield
CC pleuromutilin (PubMed:28924980). {ECO:0000250|UniProtKB:A0A2L0VXR0,
CC ECO:0000269|PubMed:28924980}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:28924980};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000250|UniProtKB:Q40577};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000269|PubMed:28924980}.
CC -!- DOMAIN: the class II DTC active site contains a DXDD motif from which
CC the middle Asp acts as the catalytic acid in this protonation-initiated
CC cyclization reaction, while the class I DTS active site contains a
CC DDXXD motif that binds divalent magnesium cofactors required for the
CC catalyzed allylic diphosphate ester ionization-initiated reaction, with
CC the first Asp playing a particularly key role.
CC {ECO:0000269|PubMed:28924980}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; LC314149; BCI98771.1; -; Genomic_DNA.
DR SMR; A0A6S6QR11; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0016829; F:lyase 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; IPR008930; Terpenoid_cyclase/PrenylTrfase.
DR SUPFAM; SSF48239; SSF48239; 1.
PE 1: Evidence at protein level;
KW Isomerase; Lyase; Magnesium; Metal-binding.
FT CHAIN 1..959
FT /note="Bifunctional premutilin synthase"
FT /id="PRO_0000453724"
FT REGION 1..542
FT /note="Class II diterpene cyclase"
FT /evidence="ECO:0000250|UniProtKB:A0A2L0VXR0"
FT REGION 543..959
FT /note="Class I diterpene synthase"
FT /evidence="ECO:0000250|UniProtKB:A0A2L0VXR0"
FT REGION 931..959
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT MOTIF 309..312
FT /note="DXDD motif"
FT /evidence="ECO:0000250|UniProtKB:A0A2L0VXR0"
FT MOTIF 649..653
FT /note="DDXXD motif"
FT /evidence="ECO:0000250|UniProtKB:A0A2L0VXR0"
FT COMPBIAS 936..959
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 311
FT /note="For class II diterpene cyclase activity"
FT /evidence="ECO:0000250|UniProtKB:A0A2L0VXR0"
FT ACT_SITE 649
FT /note="For class I diterpene synthase activity"
FT /evidence="ECO:0000250|UniProtKB:A0A2L0VXR0"
FT BINDING 649
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 649
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 653
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 653
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 824
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
SQ SEQUENCE 959 AA; 108110 MW; B8A437C66F21CFD4 CRC64;
MGLSEDLHAR ARTLMQTLES ALNTPGSRGI GTANPTIYDT AWVAMVSREI DGKQVFVFPE
TFTYIYEHQE ADGSWSGDGS LIDSIVNTLA CLLALKMHES NASKPDIPAR ARAAQHYLND
ALQRWDIMET ERVAYEMIVP CLLKQLDTFG VSFTFPHRDL LYNMYAGKLA KLNWEAIYAK
NSSLLHCMEA FVGVCDFDRM PHLLRDGNFM ATPSTTAAYL MKATKWDDRA ENYLRHVIEV
YAPHGRDVVP NLWPMTFFEI VWSLSSLYDN NLDFAQMDPE CLDRIALKLR EFLVAGKGVL
GFVPGTTHDA DMSSKTLMLL QVLNHPYSHD EFVTEFEAPT YFRCYSFERN ASVTVNSNCL
MSLLHAPDVN KYESQIVKIA TYVADVWWTS AGVVKDKWNV SEWYSSMLSS QALVRLLFEH
GKGNLKSISE ELLSKVSIAC FTMISRILQS QKPDGSWGCA EETSYALITL ANVASLPTCD
LIRDHLNQVI ESAKAFLTSI FYARPAAKPE DRVWIDKVTY SVESFRDAYL VSALNVPIPR
FDPASITTLP PISQTLPKEL AKFFGRLDMF KPAPDWRKLT WGIEATLMGP ELNRVPSLTF
EKVEKGANGK WFEFLPYMTI APSGLQGTPI SSQGMLDVLV LIRALYNTDD YLDMTLIKAT
NKDLNDLKKK IRDLFANPTS FSTLNEVPDD RMPTHIEVIE RFAHSLLNYP RIQLASDNDK
NLLRSEIEHY FLAGIAQCEE NILLRERGLD KERVGTSHYR WTHVIGADNV AGTIALVFAF
CLLGHQINEE RGSRDLVDVF PTPVLKYLFN DCVMHFGAFS RLANDLHSIS RDFNEVNLNS
IMFSEFTGPK SGTDTEKARE AALLELVKYE RKETDDAFEY LVQQLTPHVG AKRARDYINI
IRVTYLHTAL YDDLGRLTRA DISNANREVS KGANGVKKTN GLTTNGTKAT ANGSNGIHH
