ASCE_ACREG
ID ASCE_ACREG Reviewed; 1064 AA.
AC A0A455R5H4;
DT 26-FEB-2020, integrated into UniProtKB/Swiss-Prot.
DT 05-JUN-2019, sequence version 1.
DT 03-AUG-2022, entry version 11.
DE RecName: Full=Bifunctional cytochrome P450/NADPH--P450 reductase ascE {ECO:0000303|PubMed:30952781};
DE EC=1.14.14.- {ECO:0000269|PubMed:30952781};
DE AltName: Full=Ascofuranone/ascochlorin biosynthesis clusters protein E {ECO:0000303|PubMed:30952781};
GN Name=ascE {ECO:0000303|PubMed:30952781};
OS Acremonium egyptiacum (Oospora egyptiaca).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Hypocreales incertae sedis; Acremonium.
OX NCBI_TaxID=749675;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, DISRUPTION
RP PHENOTYPE, INDUCTION, AND PATHWAY.
RC STRAIN=F-1392;
RX PubMed=30952781; DOI=10.1073/pnas.1819254116;
RA Araki Y., Awakawa T., Matsuzaki M., Cho R., Matsuda Y., Hoshino S.,
RA Shinohara Y., Yamamoto M., Kido Y., Inaoka D.K., Nagamune K., Ito K.,
RA Abe I., Kita K.;
RT "Complete biosynthetic pathways of ascofuranone and ascochlorin in
RT Acremonium egyptiacum.";
RL Proc. Natl. Acad. Sci. U.S.A. 116:8269-8274(2019).
RN [2]
RP BIOTECHNOLOGY.
RX PubMed=12084465; DOI=10.1016/s0925-4439(02)00086-8;
RA Nihei C., Fukai Y., Kita K.;
RT "Trypanosome alternative oxidase as a target of chemotherapy.";
RL Biochim. Biophys. Acta 1587:234-239(2002).
RN [3]
RP BIOTECHNOLOGY.
RX PubMed=12798927; DOI=10.1016/s1383-5769(03)00012-6;
RA Yabu Y., Yoshida A., Suzuki T., Nihei C., Kawai K., Minagawa N.,
RA Hosokawa T., Nagai K., Kita K., Ohta N.;
RT "The efficacy of ascofuranone in a consecutive treatment on Trypanosoma
RT brucei brucei in mice.";
RL Parasitol. Int. 52:155-164(2003).
RN [4]
RP BIOTECHNOLOGY.
RX PubMed=20688188; DOI=10.1016/j.parint.2010.07.006;
RA Nakamura K., Fujioka S., Fukumoto S., Inoue N., Sakamoto K., Hirata H.,
RA Kido Y., Yabu Y., Suzuki T., Watanabe Y., Saimoto H., Akiyama H., Kita K.;
RT "Trypanosome alternative oxidase, a potential therapeutic target for
RT sleeping sickness, is conserved among Trypanosoma brucei subspecies.";
RL Parasitol. Int. 59:560-564(2010).
CC -!- FUNCTION: Bifunctional cytochrome P450/NADPH--P450 reductase; part of
CC the asc-1 gene cluster that mediates the biosynthesis both ascochlorin
CC and ascofuranone, a strong inhibitor of cyanide-insensitive alternative
CC oxidases and a promising drug candidate against African trypanosomiasis
CC (PubMed:30952781). The first step in the pathway is performed by the
CC non-reducing polyketide synthase ascC that produces orsellinic acid by
CC condensing acetyl-CoA with 3 malonyl-CoA units (PubMed:30952781).
CC Orsellinic acid is then prenylated by the prenyltransferase ascA to
CC yield ilicicolinic acid B (PubMed:30952781). Ilicicolinic acid B is
CC further reduced to ilicicolin B by the reductase ascB
CC (PubMed:30952781). The halogenase ascD then chlorinates ilicicolin B to
CC produce ilicicolin A which is converted to ilicicolin A epoxide by the
CC cytochrome P450 monooxygenase ascE that catalyzes stereoselective
CC epoxidation of the terminal double bond of the prenyl group
CC (PubMed:30952781). Ilicicolin A epoxide is the last common precursor
CC for the biosynthesis of ascofuranone and ascochlorin (PubMed:30952781).
CC The terpene cyclase ascF produces a monocyclic terpene, and the
CC cyclization reaction is proposed to be initiated by protonation of the
CC terminal epoxide of ilicicolin A epoxide to generate a monocyclic
CC tertiarycation, which is followed by a series of hydride and methyl
CC shifts with abstraction of proton, leading to the formation of the
CC (14S,15R,19R)-trimethylcyclohexanone ring structure of ilicicolin C,
CC which is finally reduced to ascochlorin by the dehydrogenase ascG
CC (PubMed:30952781). On the other hand, ilicicolin A epoxide is
CC hydroxylated by the cytochrome P450 monooxygenase ascH, and the
CC resultant product is cyclized by the terpene cyclase ascI to
CC ascofuranol via protonation-initiated epoxide ring opening, which
CC facilitates the 6-endo-tet cyclization to form the tetrahy-drofuran
CC ring (PubMed:30952781). Finally, ascofuranol is oxidized into
CC ascofuranone by ascJ (PubMed:30952781). {ECO:0000269|PubMed:30952781}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + ilicicolin A + NADPH + O2 = H2O + ilicicolin A epoxide
CC + NADP(+); Xref=Rhea:RHEA:63092, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:146154, ChEBI:CHEBI:146155;
CC Evidence={ECO:0000269|PubMed:30952781};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:63093;
CC Evidence={ECO:0000269|PubMed:30952781};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC Note=Binds 1 FAD. {ECO:0000250|UniProtKB:Q9Y8G7};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC Note=Binds 1 FMN. {ECO:0000250|UniProtKB:Q9Y8G7};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000269|PubMed:30952781}.
CC -!- INDUCTION: Expression is induced on AF medium.
CC {ECO:0000269|PubMed:30952781}.
CC -!- DISRUPTION PHENOTYPE: Impairs the production of ascochlorin and
CC ascofuranone, and leads to the accumulation of ilicicolin A.
CC {ECO:0000269|PubMed:30952781}.
CC -!- BIOTECHNOLOGY: Ascofuranone is a specific inhibitor of trypanosome
CC alternative oxidase (TAO), and quickly kills African trypanosomes in
CC vitro and cures infected mice. As an essential factor for trypanosome
CC survival, TAO is a promising drug target due to the absence of
CC alternative oxidases in the mammalian host.
CC {ECO:0000269|PubMed:12084465, ECO:0000269|PubMed:12798927,
CC ECO:0000269|PubMed:20688188}.
CC -!- SIMILARITY: In the N-terminal section; belongs to the cytochrome P450
CC family. {ECO:0000305}.
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DR EMBL; LC406756; BBF25317.1; -; Genomic_DNA.
DR AlphaFoldDB; A0A455R5H4; -.
DR SMR; A0A455R5H4; -.
DR UniPathway; UPA00213; -.
DR GO; GO:0070330; F:aromatase activity; IEA:InterPro.
DR GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0003958; F:NADPH-hemoprotein reductase activity; IEA:InterPro.
DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 1.10.630.10; -; 1.
DR Gene3D; 1.20.990.10; -; 1.
DR Gene3D; 3.40.50.360; -; 1.
DR Gene3D; 3.40.50.80; -; 1.
DR InterPro; IPR023206; Bifunctional_P450_P450_red.
DR InterPro; IPR003097; CysJ-like_FAD-binding.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR InterPro; IPR017927; FAD-bd_FR_type.
DR InterPro; IPR008254; Flavodoxin/NO_synth.
DR InterPro; IPR029039; Flavoprotein-like_sf.
DR InterPro; IPR039261; FNR_nucleotide-bd.
DR InterPro; IPR023173; NADPH_Cyt_P450_Rdtase_alpha.
DR InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR Pfam; PF00667; FAD_binding_1; 1.
DR Pfam; PF00258; Flavodoxin_1; 1.
DR Pfam; PF00175; NAD_binding_1; 1.
DR Pfam; PF00067; p450; 1.
DR PIRSF; PIRSF000209; Bifunctional_P450_P450R; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR SUPFAM; SSF52218; SSF52218; 1.
DR SUPFAM; SSF52343; SSF52343; 1.
DR SUPFAM; SSF63380; SSF63380; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
DR PROSITE; PS51384; FAD_FR; 1.
DR PROSITE; PS50902; FLAVODOXIN_LIKE; 1.
PE 1: Evidence at protein level;
KW Electron transport; FAD; Flavoprotein; FMN; Heme; Iron; Metal-binding;
KW Monooxygenase; NADP; Oxidoreductase; Transport.
FT CHAIN 1..1064
FT /note="Bifunctional cytochrome P450/NADPH--P450 reductase
FT ascE"
FT /id="PRO_0000448999"
FT DOMAIN 504..644
FT /note="Flavodoxin-like"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT DOMAIN 676..905
FT /note="FAD-binding FR-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT REGION 1..484
FT /note="Cytochrome P450"
FT /evidence="ECO:0000250|UniProtKB:Q9Y8G7"
FT REGION 485..1064
FT /note="NADPH-P-450 reductase"
FT /evidence="ECO:0000250|UniProtKB:Q9Y8G7"
FT BINDING 411
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250|UniProtKB:P14779"
FT BINDING 510..514
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT BINDING 588..620
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT BINDING 591
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000250|UniProtKB:P14779"
FT BINDING 599
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000250|UniProtKB:P14779"
FT SITE 271
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000250|UniProtKB:P14779"
SQ SEQUENCE 1064 AA; 117894 MW; 8D004CDA96C30ABB CRC64;
MTELIPGPKG LPLIGNVLDI DPVDAVVCLG RIADTYGHIY QLKVGGSAKI FISSRELVDE
LSDESRFTKL VSGPLAQLRN VCHDSLFTAQ SDEPAWDLAH KILMPAFGPL AIRGMFDEMH
DIASQLVVKW ARFGPQDTID VSGDFTRLTL DAIALCSMST RFNSFYKQDQ HPFVSSMLEV
LAESGKRAVR PPFVNDYIFR GSLKHYNTEI ATMRRIAMDV LAERRANPMA CQKNDLLNAM
INGRDPKTGE GLSDESTINN LIVFLIAGHE TTSGLLSFLF YYLLTRPDVF EKAQKEVDEL
VGRGPVTIEH MSKLHYIEAC LRETLRLHPT APVITFKTKP GFEKESTTIG GGKYKIDRDQ
GIVALLVNIQ RDPKVWGDDA NEFKPERMTD EKFNNLPANC WKPFGNGIRG CIGRAFAWQE
SLLITAMLLQ NFNFQLADPD YKLQIKQTLT IKPGNFFMHA KLRDHVDPLE LEGILHGGAK
KGSKIDGPSS GASLATTEQE LQPMTILYGS DSGTCESMAQ SLARAARGRG YGATVKTLDS
AVEQVPKDQP VVIVSPSYNG QPPSNATDFV KWLEALDSKA LKDVKYSVYG CGNKDYTSTF
HRIPKLLDAE FERCGAKRIA ETGLGDVTVG DIFSDFERWQ DDQLWPALGV AHMDGDADAE
FDIHVDRSGR AAELEVDADE ATVQSNQVLT APGEPEKRYI TLKLPEGMQY KSGDHLSVLP
LNDWGVVRRV FAWAQLPWDA VVTIPKGTNT SLPTGRQISA KDLLSGYVEL SQPATRKNIA
KLAASSPCPF TQKSLSKLEE HFDSDIAQRR LSVLDILEEF PAIDITFGNF ISMLPPMRPR
QYSIASSPMA DPSTATLMWT VLNSEAYSGS GRRFLGVCST YLAGLAEGDR VHVTVKPALR
LFHPPSDPES MPIIMACAGT GLAPFRGFLE ERVCQMKAGR ALAPAYLFVG CRDPEKDALL
KDELAQWERD GVVKIYYAFS RASDQSDGCK HVQDRIWNER DLVRKGLFEG NARFFMCGGS
GAGKSVEDVV KRIYKDNKGE SQEKAAESWF QDLKANRYVT EIFA
