MPAG_PENBR
ID MPAG_PENBR Reviewed; 398 AA.
AC F1DBB3;
DT 06-JUL-2016, integrated into UniProtKB/Swiss-Prot.
DT 03-MAY-2011, sequence version 1.
DT 03-AUG-2022, entry version 41.
DE RecName: Full=O-methyltransferase mpaG {ECO:0000303|PubMed:21398490};
DE EC=2.1.1.- {ECO:0000269|PubMed:25630520};
DE AltName: Full=Mycophenolic acid biosynthesis cluster protein G {ECO:0000303|PubMed:21398490};
GN Name=mpaG {ECO:0000303|PubMed:21398490};
OS Penicillium brevicompactum.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=5074;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=IBT 23078;
RX PubMed=21398490; DOI=10.1128/aem.03015-10;
RA Regueira T.B., Kildegaard K.R., Hansen B.G., Mortensen U.H., Hertweck C.,
RA Nielsen J.;
RT "Molecular basis for mycophenolic acid biosynthesis in Penicillium
RT brevicompactum.";
RL Appl. Environ. Microbiol. 77:3035-3043(2011).
RN [2]
RP FUNCTION.
RC STRAIN=IBT23078;
RX PubMed=22544261; DOI=10.1128/aem.07955-11;
RA Hansen B.G., Mnich E., Nielsen K.F., Nielsen J.B., Nielsen M.T.,
RA Mortensen U.H., Larsen T.O., Patil K.R.;
RT "Involvement of a natural fusion of a cytochrome p450 and a hydrolase in
RT mycophenolic acid biosynthesis.";
RL Appl. Environ. Microbiol. 78:4908-4913(2012).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, MUTAGENESIS OF HIS-306; GLU-335 AND GLU-362,
RP ACTIVE SITE, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX PubMed=25630520; DOI=10.1002/cbic.201402600;
RA Zhang W., Cao S., Qiu L., Qi F., Li Z., Yang Y., Huang S., Bai F., Liu C.,
RA Wan X., Li S.;
RT "Functional characterization of MpaG', the O-methyltransferase involved in
RT the biosynthesis of mycophenolic acid.";
RL ChemBioChem 16:565-569(2015).
CC -!- FUNCTION: O-methyltransferase; part of the gene cluster that mediates
CC the biosynthesis of mycophenolic acid (MPA), the first isolated
CC antibiotic natural product in the world obtained from a culture of
CC Penicillium brevicompactum in 1893 (PubMed:25630520). MpaC methylates
CC farnesyl-DHMP-3C (FDHMP-3C) to yield MFDHMP-3C (PubMed:25630520). The
CC first step of the pathway is the synthesis of 5-methylorsellinic acid
CC (5MOA) by the cytosolic polyketide synthase mpaC. 5MOA is then
CC converted to the phthalide compound 5,7-dihydroxy-4,6-dimethylphthalide
CC (DHMP) by the endoplasmic reticulum-bound cytochrome P450 monooxygenase
CC mpaDE. MpaDE first catalyzes hydroxylation of 5-MOA to 4,6-dihydroxy-2-
CC (hydroxymethyl)-3-methylbenzoic acid (DHMB). MpaDE then acts as a
CC lactone synthase that catalyzes the ring closure to convert DHMB into
CC DHMP. The next step is the prenylation of DHMP by the Golgi apparatus-
CC associated prenyltransferase mpaA to yield farnesyl-DHMP (FDHMP). The
CC ER-bound oxygenase mpaB then mediates the oxidative cleavage the C19-
CC C20 double bond in FDHMP to yield FDHMP-3C via a mycophenolic aldehyde
CC intermediate. The O-methyltransferase mpaG catalyzes the methylation of
CC FDHMP-3C to yield MFDHMP-3C. After the cytosolic methylation of FDHMP-
CC 3C, MFDHMP-3C enters into peroxisomes probably via free diffusion due
CC to its low molecular weight. Upon a peroxisomal CoA ligation reaction,
CC catalyzed by a beta-oxidation component enzyme acyl-CoA ligase ACL891,
CC MFDHMP-3C-CoA would then be restricted to peroxisomes for the following
CC beta-oxidation pathway steps. The peroxisomal beta-oxidation machinery
CC than converts MFDHMP-3C-CoA into MPA_CoA, via a beta-oxidation chain-
CC shortening process. Finally mpaH acts as a peroxisomal acyl-CoA
CC hydrolase with high substrate specificity toward MPA-CoA to release the
CC final product MPA (PubMed:21398490, PubMed:22544261) (Probable).
CC {ECO:0000269|PubMed:25630520, ECO:0000305|PubMed:21398490,
CC ECO:0000305|PubMed:22544261}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(4E,8E)-10-(4,6-dihydroxy-7-methyl-3-oxo-1,3-dihydro-2-
CC benzofuran-5-yl)-4,8-dimethyldeca-4,8-dienoate + S-adenosyl-L-
CC methionine = (4E,8E)-10-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1,3-
CC dihydro-2-benzofuran-5-yl)-4,8-dimethyldeca-4,8-dienoate + H(+) + S-
CC adenosyl-L-homocysteine; Xref=Rhea:RHEA:66696, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57856, ChEBI:CHEBI:59789, ChEBI:CHEBI:167389,
CC ChEBI:CHEBI:167390; Evidence={ECO:0000269|PubMed:25630520};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66697;
CC Evidence={ECO:0000269|PubMed:25630520};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=51.5 uM for demethylmycophenolic acid (DMMPA)
CC {ECO:0000269|PubMed:25630520};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:25630520};
CC Temperature dependence:
CC Optimum temperature is 45 degrees Celsius.
CC {ECO:0000269|PubMed:25630520};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000269|PubMed:25630520}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol
CC {ECO:0000250|UniProtKB:A0A0B5L781}.
CC -!- SIMILARITY: Belongs to the class I-like SAM-binding methyltransferase
CC superfamily. Cation-independent O-methyltransferase family. COMT
CC subfamily. {ECO:0000305}.
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DR EMBL; HQ731031; ADY00134.1; -; Genomic_DNA.
DR AlphaFoldDB; F1DBB3; -.
DR SMR; F1DBB3; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0005829; C:cytosol; ISS:GO_Central.
DR GO; GO:0008168; F:methyltransferase activity; ISS:GO_Central.
DR GO; GO:0008171; F:O-methyltransferase activity; IEA:InterPro.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR GO; GO:0140722; P:mycophenolic acid biosynthetic process; ISS:GO_Central.
DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 1.10.10.10; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR InterPro; IPR016461; COMT-like.
DR InterPro; IPR001077; O_MeTrfase_dom.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR InterPro; IPR036388; WH-like_DNA-bd_sf.
DR InterPro; IPR036390; WH_DNA-bd_sf.
DR Pfam; PF00891; Methyltransf_2; 1.
DR PIRSF; PIRSF005739; O-mtase; 1.
DR SUPFAM; SSF46785; SSF46785; 1.
DR SUPFAM; SSF53335; SSF53335; 1.
DR PROSITE; PS51683; SAM_OMT_II; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Methyltransferase; S-adenosyl-L-methionine; Transferase.
FT CHAIN 1..398
FT /note="O-methyltransferase mpaG"
FT /id="PRO_0000436575"
FT ACT_SITE 306
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01020"
FT ACT_SITE 335
FT /evidence="ECO:0000269|PubMed:25630520"
FT ACT_SITE 362
FT /evidence="ECO:0000269|PubMed:25630520"
FT BINDING 264
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01020"
FT MUTAGEN 306
FT /note="H->A: Completely looses methyltransferase activity
FT towards demethylmycophenolic acid (DMMPA)."
FT /evidence="ECO:0000269|PubMed:25630520"
FT MUTAGEN 335
FT /note="E->A: Completely looses methyltransferase activity
FT towards demethylmycophenolic acid (DMMPA)."
FT /evidence="ECO:0000269|PubMed:25630520"
FT MUTAGEN 362
FT /note="E->A: Completely looses methyltransferase activity
FT towards demethylmycophenolic acid (DMMPA)."
FT /evidence="ECO:0000269|PubMed:25630520"
SQ SEQUENCE 398 AA; 43434 MW; CE40674483BC1687 CRC64;
MSAASPASII QELASAAKQY ENNESGAREA LIAQSRALIA SLEVPSEFIQ HTFWSQPALS
AIVRLATDVN LFQYLKDAQE EGLSAEALAS KTGMDVSLFA RLARHLVAMN VITSRNGVFY
GTALSNGLAA ENYQQSIRFC HDVSRPSFGA FPSFFKGNGY KTPALGTTDG PFQSAHKVDI
SFPQWLVGNP PYLQYFNSYM SAYRAGKPNW CDNGFYPVAD RLLNGFDASV SDVLLVDVGG
GRGHDIATFG SQFSPLPGRL VLQDREQVIN SIPADESRQF EATTHDIFTT QPVKNARAYY
MHSVPHGFGD EDAVKIMANL VPALAKGYSR VLLNEIVVDE ESPVMSATNM DLIMLAHMGA
KERTEADWRS ILTRAGLKVV NIYSYPGVAE SLIEAELA
