CPAD_ASPOZ
ID CPAD_ASPOZ Reviewed; 437 AA.
AC F5HN73;
DT 10-OCT-2018, integrated into UniProtKB/Swiss-Prot.
DT 27-JUL-2011, sequence version 1.
DT 03-AUG-2022, entry version 39.
DE RecName: Full=Cyclo-acetoacetyl-L-tryptophan dimethylallyltransferase cpaD {ECO:0000303|PubMed:19877600};
DE EC=2.5.1.- {ECO:0000269|PubMed:19877600};
DE AltName: Full=Cyclopiazonic acid biosynthesis cluster protein D {ECO:0000303|PubMed:21608094};
DE AltName: Full=L-tryptophan dimethylallyl transferase {ECO:0000305};
DE Short=DMATS {ECO:0000305};
GN Name=cpaD {ECO:0000303|PubMed:21608094};
OS Aspergillus oryzae (Yellow koji mold).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=5062;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], DISRUPTION PHENOTYPE, FUNCTION, AND
RP PATHWAY.
RC STRAIN=NBRC 4177;
RX PubMed=21608094; DOI=10.1002/cbic.201000672;
RA Kato N., Tokuoka M., Shinohara Y., Kawatani M., Uramoto M., Seshime Y.,
RA Fujii I., Kitamoto K., Takahashi T., Takahashi S., Koyama Y., Osada H.;
RT "Genetic safeguard against mycotoxin cyclopiazonic acid production in
RT Aspergillus oryzae.";
RL ChemBioChem 12:1376-1382(2011).
RN [2]
RP FUNCTION.
RX PubMed=19663400; DOI=10.1021/bi901123r;
RA Liu X., Walsh C.T.;
RT "Cyclopiazonic acid biosynthesis in Aspergillus sp.: characterization of a
RT reductase-like R* domain in cyclopiazonate synthetase that forms and
RT releases cyclo-acetoacetyl-L-tryptophan.";
RL Biochemistry 48:8746-8757(2009).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX PubMed=19877600; DOI=10.1021/bi901597j;
RA Liu X., Walsh C.T.;
RT "Characterization of cyclo-acetoacetyl-L-tryptophan
RT dimethylallyltransferase in cyclopiazonic acid biosynthesis: substrate
RT promiscuity and site directed mutagenesis studies.";
RL Biochemistry 48:11032-11044(2009).
CC -!- FUNCTION: Cyclo-acetoacetyl-L-tryptophan dimethylallyltransferase; part
CC of the gene cluster that mediates the biosynthesis of the fungal
CC neurotoxin cyclopiazonic acid (CPA), a nanomolar inhibitor of Ca(2+)-
CC ATPase with a unique pentacyclic indole tetramic acid scaffold
CC (PubMed:21608094). The hybrid two module polyketide synthase-
CC nonribosomal peptide synthetase (PKS-NRPS) cpaS incorporates acetyl-
CC CoA, malonyl-CoA, and tryptophan (Trp) and utilizes a C-terminal redox-
CC incompetent reductase domain to make and release the tryptophan
CC tetramic acid, cyclo-acetoacetyl-L-tryptophan (c-AATrp), as the first
CC intermediate in the pathway. CpaS catalyzes a Dieckmann-type
CC cyclization on the N-acetoacetyl-Trp intermediate bound in thioester
CC linkage to the phosphopantetheinyl arm of the T domain to form and
CC release c-AATrp (PubMed:21608094, PubMed:19663400). CpaD than
CC regiospecifically dimethylallylates c-AATrp to form beta-cyclopiazonic
CC acid. CpaD discriminates against free Trp but accepts tryptophan-
CC containing thiohydantoins, diketopiperazines, and linear peptides as
CC substrates for C4-prenylation and also acts as regiospecific O-
CC dimethylallyltransferase (DMAT) on a tyrosine-derived tetramic acid
CC (PubMed:21608094, PubMed:19877600). The beta-cyclopiazonate
CC dehydrogenase cpaO then carries out the dehydrogenation of beta-CPA to
CC yield an unstable enimine product, which is captured by intramolecular
CC cyclization to create the pentacyclic fused scaffold of alpha-
CC cyclopiazonate (PubMed:21608094). Fimally, the cytochrome P450
CC monooxygenase cpaH mediates the conversion of CPA into the less toxic
CC 2-oxocyclopiazonic acid, the end product of the CPA pathway in A.oryza
CC (PubMed:21608094). {ECO:0000269|PubMed:19663400,
CC ECO:0000269|PubMed:19877600, ECO:0000269|PubMed:21608094}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cyclo-acetoacetyl-L-tryptophan + dimethylallyl diphosphate =
CC beta-cyclopiazonate + diphosphate; Xref=Rhea:RHEA:10384,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57623, ChEBI:CHEBI:58067,
CC ChEBI:CHEBI:167552; Evidence={ECO:0000269|PubMed:19877600};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:10385;
CC Evidence={ECO:0000269|PubMed:19877600};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=109 uM for L-c-AATrp {ECO:0000269|PubMed:19877600};
CC KM=3829 uM for D-c-AATrp {ECO:0000269|PubMed:19877600};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:19877600, ECO:0000269|PubMed:21608094}.
CC -!- DISRUPTION PHENOTYPE: Impairs the production of cyclopiazonic acid and
CC of its biosynthetic intermediate beta-cyclopiazonic acid, but
CC accumulates the intermediate cyclo-acetoacetyl-L-tryptophan.
CC {ECO:0000269|PubMed:21608094}.
CC -!- SIMILARITY: Belongs to the tryptophan dimethylallyltransferase family.
CC {ECO:0000305}.
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DR EMBL; AB506492; BAK26561.1; -; Genomic_DNA.
DR AlphaFoldDB; F5HN73; -.
DR SMR; F5HN73; -.
DR VEuPathDB; FungiDB:AO090026000002; -.
DR OMA; YIYPRIK; -.
DR BioCyc; MetaCyc:MON-18881; -.
DR GO; GO:0004659; F:prenyltransferase activity; IEA:UniProt.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0044249; P:cellular biosynthetic process; IEA:UniProt.
DR GO; GO:1901576; P:organic substance biosynthetic process; IEA:UniProt.
DR CDD; cd13929; PT-DMATS_CymD; 1.
DR InterPro; IPR033964; Aro_prenylTrfase.
DR InterPro; IPR017795; Aro_prenylTrfase_DMATS.
DR InterPro; IPR012148; DMATS-type_fun.
DR PANTHER; PTHR40627; PTHR40627; 1.
DR Pfam; PF11991; Trp_DMAT; 1.
DR PIRSF; PIRSF000509; Trp_DMAT; 1.
DR SFLD; SFLDS00036; Aromatic_Prenyltransferase; 1.
DR TIGRFAMs; TIGR03429; arom_pren_DMATS; 1.
PE 1: Evidence at protein level;
KW Alkaloid metabolism; Transferase.
FT CHAIN 1..437
FT /note="Cyclo-acetoacetyl-L-tryptophan
FT dimethylallyltransferase cpaD"
FT /id="PRO_0000445386"
FT BINDING 84..85
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 93
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 104
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 191
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 193
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 195
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 258
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 260
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 262
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 344
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 346
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 410
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 414
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
SQ SEQUENCE 437 AA; 49186 MW; 67CAB188CDDDDDA0 CRC64;
MEISKKAATL LPKPFYVLSQ ALNLSNKDHT KWWYSTAPMF ATMMAGAGYD VHAQYKFLCI
HREVIIPALG PYPEKGQPMH WKSHLTRFGL PFELSFNYSK SLLRFAFEPL GSLTGTKDDP
FNTQAIRPVL QDLKAMVPGL DLEWFDHFTK ALVVSEEEAR TLLDRDIEIP VFKTQNKLAA
DLEPSGDIVL KTYIYPRIKS IATGTPKERL MFDAIKAADK FGKVATPLAI LEEFIAERAP
TLLGHFLSCD LVKPSESRIK VYCMERQLDL ASIEGIWTLN GRRNDPETLD GLDALRELWQ
LLPVTEGLCP LPNCFYEPGT SPQEQLPFII NFTLSPKSAL PEPQIYFPAF GQNDKTIAEG
LATFFESRGW GGLAKSYPAD LASYYPDVDL QTANHLQAWI SFSYKGKKPY MSVYLHTFEA
FSAAAQEVAM CHDGHNP
