PLE3_CLIPA
ID PLE3_CLIPA Reviewed; 959 AA.
AC A0A2L0VXR0;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 25-APR-2018, sequence version 1.
DT 03-AUG-2022, entry version 11.
DE RecName: Full=Bifunctional premutilin synthase {ECO:0000303|PubMed:29388775};
DE Short=PS {ECO:0000303|PubMed:29388775};
DE Includes:
DE RecName: Full=Class II diterpene cyclase {ECO:0000303|PubMed:29388775};
DE EC=5.5.1.- {ECO:0000269|PubMed:29388775};
DE Includes:
DE RecName: Full=Class I diterpene synthase {ECO:0000303|PubMed:29388775};
DE EC=4.2.3.- {ECO:0000269|PubMed:29388775};
GN Name=PS {ECO:0000303|PubMed:29388775};
GN Synonyms=cyc {ECO:0000303|PubMed:27143514};
OS Clitopilus passeckerianus (Pleurotus passeckerianus).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes;
OC Agaricomycetidae; Agaricales; Entolomataceae; Clitopilus.
OX NCBI_TaxID=648682;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION, FUNCTION, INDUCTION, AND
RP PATHWAY.
RX PubMed=27143514; DOI=10.1038/srep25202;
RA Bailey A.M., Alberti F., Kilaru S., Collins C.M., de Mattos-Shipley K.,
RA Hartley A.J., Hayes P., Griffin A., Lazarus C.M., Cox R.J., Willis C.L.,
RA O'Dwyer K., Spence D.W., Foster G.D.;
RT "Identification and manipulation of the pleuromutilin gene cluster from
RT Clitopilus passeckerianus for increased rapid antibiotic production.";
RL Sci. Rep. 6:25202-25202(2016).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, DOMAIN, ACTIVE
RP SITE, MUTAGENESIS OF ASP-311 AND ASP-649, AND PATHWAY.
RX PubMed=29388775; DOI=10.1021/acs.orglett.8b00121;
RA Xu M., Jia M., Hong Y.J., Yin X., Tantillo D.J., Proteau P.J., Peters R.J.;
RT "Premutilin synthase: ring rearrangement by a class II diterpene cyclase.";
RL Org. Lett. 20:1200-1202(2018).
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:27143514, PubMed:29388775).
CC The geranylgeranyl diphosphate (GGPP) synthase catalyzes the first step
CC in pleuromutilin biosynthesis (PubMed:27143514, PubMed:29388775). GGPP
CC is then substrate of the premutilin synthase (PS) to yield premutilin
CC (PubMed:29388775). 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. GGPP is
CC first converted to mutildienyl-diphosphate (MPP) at the class II DTC
CC site (PubMed:29388775). MPP is subsequently further cyclized at the
CC class I DTS site, followed by a 1,5-hydride shift and addition of water
CC prior to terminating deprotonation, to yield premutilin
CC (PubMed:29388775). In addition to the aforementioned GGPP synthase and
CC bifunctional diterpene synthase, the cluster contains also three
CC cytochrome P450 monooxygenases, a short-chain alcohol dehydrogenase,
CC and an acyltransferase, involved in the conversion of premutilin to
CC pleuromutilin (PubMed:27143514, PubMed:29388775). The cytochrome P450
CC monooxygenases P450-1 and P450-2 hydroxylate premutilin at C-11 and C-
CC 3, respectively, producing 11-hydroxypremutilin and 3-hydroxypremutilin
CC (By similarity). The combination of the actions of both ple5 and ple6
CC leads to the production of 3,11-dihydroxypremutilin (By similarity).
CC The short chain dehydrogenase SDR further converts 3,11-
CC dihydroxypremutilin into mutilin (By similarity). The acetyltransferase
CC ATF then acetylates mutilin to produce 14-O-acetylmutilin (By
CC similarity). Finally, the cytochrome P450 monooxygenase P450-3
CC catalyzes hydroxylation on the alpha position of the acetyl side chain
CC of 14-O-acetylmutilin to yield pleuromutilin (By similarity).
CC {ECO:0000250|UniProtKB:A0A6S6QR11, ECO:0000269|PubMed:27143514,
CC ECO:0000269|PubMed:29388775}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:Q40577};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000250|UniProtKB:Q40577};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000269|PubMed:27143514, ECO:0000269|PubMed:29388775}.
CC -!- INDUCTION: Expression is up-regulated during pleuromutilin production.
CC {ECO:0000269|PubMed:27143514}.
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:29388775}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; MG764077; AVA16672.1; -; mRNA.
DR AlphaFoldDB; A0A2L0VXR0; -.
DR SMR; A0A2L0VXR0; -.
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:0017000; P:antibiotic biosynthetic process; 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 Antibiotic biosynthesis; Isomerase; Lyase; Magnesium; Metal-binding.
FT CHAIN 1..959
FT /note="Bifunctional premutilin synthase"
FT /id="PRO_0000445371"
FT REGION 1..542
FT /note="Class II diterpene cyclase"
FT /evidence="ECO:0000305|PubMed:29388775"
FT REGION 543..959
FT /note="Class I diterpene synthase"
FT /evidence="ECO:0000305|PubMed:29388775"
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:0000305|PubMed:29388775"
FT MOTIF 649..653
FT /note="DDXXD motif"
FT /evidence="ECO:0000305|PubMed:29388775"
FT ACT_SITE 311
FT /note="For class II diterpene cyclase activity"
FT /evidence="ECO:0000269|PubMed:29388775"
FT ACT_SITE 649
FT /note="For class I diterpene synthase activity"
FT /evidence="ECO:0000269|PubMed:29388775"
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"
FT MUTAGEN 311
FT /note="D->A: Blocks class II DTC activity and abolishes the
FT production of premutilin."
FT /evidence="ECO:0000269|PubMed:29388775"
FT MUTAGEN 649
FT /note="D->L: Completely blocks class I DTS activity,
FT abolishes the production of premutilin, and leads to the
FT accumulation of mutildienyl-diphosphate."
FT /evidence="ECO:0000269|PubMed:29388775"
SQ SEQUENCE 959 AA; 107841 MW; A1BC6B9AF3338686 CRC64;
MGLSEDLHAR ARTLMQTLES ALNTPGSRGI GTANPTIYDT AWVAMVSREI DGKQVFVFPE
TFTYIYEHQE ADGSWSGDGS LIDSIVNTLA CLVALKMHES NASKPDIPAR ARAAQNYLDD
ALKRWDIMET ERVAYEMIVP CLLKQLDAFG VSFTFPHHDL LYNMYAGKLA KLNWEAIYAK
NSSLLHCMEA FVGVCDFDRM PHLLRDGNFM ATPSTTAAYL MKATKWDDRA EDYLRHVIEV
YAPHGRDVVP NLWPMTFFEI VWSLSSLYDN NLEFAQMDPE CLDRIALKLR EFLVAGKGVL
GFVPGTTHDA DMSSKTLMLL QVLNHPYSHD EFVTEFEAPT YFRCYSFERN ASVTVNSNCL
MSLLHAPDVN KYESQIVKIA TYVADVWWTS AGVVKDKWNV SEWYSSMLSS QALVRLLFEH
GKGNLKSISE ELLSRVSIAC FTMISRILQS QKPDGSWGCA EETSYALITL ANVASLPTCD
LIRDHLYKVI ESAKAYLTPI FYARPAAKPE DRVWIDKVTY SVESFRDAYL VSALNVPIPR
FDPSSISTLP AISQTLPKEL SKFFGRLDMF KPAPEWRKLT WGIEATLMGP ELNRVPSSTF
AKVEKGAAGK WFEFLPYMTI APSSLEGTPI SSQGMLDVLV LIRGLYNTDD YLDMTLIKAT
NEDLDDLKKK IRDLFADPKS FSTLSEVPDD RMPTHIEVIE RFAYSLLNHP RAQLASDNDK
GLLRSEIEHY FLAGIAQCEE NILLRERGLD KERIGTSHYR WTHVVGADNV AGTIALVFAL
CLLGHQINEE RGSRDLVDVF PSPVLKYLFN DCVMHFGTFS RLANDLHSIS RDFNEVNLNS
IMFSEFTGPK SGTDTEKARE AALLELTKFE RKATDDGFEY LVQQLTPHVG AKRARDYINI
IRVTYLHTAL YDDLGRLTRA DISNANQEVS KGTNGVKKIN GSSTNGTKVT ANGSNGIHH
