5MOAR_ASPTN
ID 5MOAR_ASPTN Reviewed; 1069 AA.
AC Q0CRQ4;
DT 17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT 17-OCT-2006, sequence version 1.
DT 03-AUG-2022, entry version 79.
DE RecName: Full=Adenylate-forming reductase {ECO:0000305};
DE EC=1.2.1.- {ECO:0000269|PubMed:24412543};
DE AltName: Full=Azasperpyranone A biosynthesis cluster A protein ATEG_03630 {ECO:0000303|PubMed:31908094};
DE AltName: Full=Nonribosomal peptide synthase-like protein ATEG_03630 {ECO:0000303|PubMed:24412543};
DE Short=NRPS-like protein ATEG_03630 {ECO:0000303|PubMed:24412543};
GN ORFNames=ATEG_03630;
OS Aspergillus terreus (strain NIH 2624 / FGSC A1156).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=341663;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=NIH 2624 / FGSC A1156;
RA Birren B.W., Lander E.S., Galagan J.E., Nusbaum C., Devon K., Henn M.,
RA Ma L.-J., Jaffe D.B., Butler J., Alvarez P., Gnerre S., Grabherr M.,
RA Kleber M., Mauceli E.W., Brockman W., Rounsley S., Young S.K., LaButti K.,
RA Pushparaj V., DeCaprio D., Crawford M., Koehrsen M., Engels R.,
RA Montgomery P., Pearson M., Howarth C., Larson L., Luoma S., White J.,
RA Alvarado L., Kodira C.D., Zeng Q., Oleary S., Yandava C., Denning D.W.,
RA Nierman W.C., Milne T., Madden K.;
RT "Annotation of the Aspergillus terreus NIH2624 genome.";
RL Submitted (SEP-2005) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION.
RX PubMed=23621425; DOI=10.1021/ja401945a;
RA Chiang Y.M., Oakley C.E., Ahuja M., Entwistle R., Schultz A., Chang S.L.,
RA Sung C.T., Wang C.C., Oakley B.R.;
RT "An efficient system for heterologous expression of secondary metabolite
RT genes in Aspergillus nidulans.";
RL J. Am. Chem. Soc. 135:7720-7731(2013).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, DOMAIN, MUTAGENESIS OF SER-612; THR-690;
RP TYR-863 AND LYS-867, AND PATHWAY.
RX PubMed=24412543; DOI=10.1016/j.chembiol.2013.12.005;
RA Wang M., Beissner M., Zhao H.;
RT "Aryl-aldehyde formation in fungal polyketides: discovery and
RT characterization of a distinct biosynthetic mechanism.";
RL Chem. Biol. 21:257-263(2014).
RN [4]
RP FUNCTION, INDUCTION, DISRUPTION PHENOTYPE, CATALYTIC ACTIVITY, PATHWAY, AND
RP BIOTECHNOLOGY.
RX PubMed=31908094; DOI=10.1002/anie.201915514;
RA Huang X., Zhang W., Tang S., Wei S., Lu X.;
RT "Collaborative biosynthesis of a class of bioactive azaphilones by two
RT separate gene clusters containing four PKS/NRPSs with transcriptional
RT cosstalk in fungi.";
RL Angew. Chem. Int. Ed. 59:4349-4353(2020).
CC -!- FUNCTION: Non-canonical non-ribosomal peptide synthetase; part of the
CC cluster A that mediates the biosynthesis of azasperpyranones, members
CC of the azaphilone family that exhibit anti-cancer activities
CC (PubMed:31908094). Azasperpyranones are synthesized by 2 clusters, A
CC and B (PubMed:31908094). Cluster A is responsible for the production of
CC the polyhydric phenol moiety while the azaphilonoid scaffold is
CC produced by the cluster B (PubMed:31908094). The non-reducing
CC polyketide synthase ATEG_03629 produces 5-methyl orsellinic acid, which
CC is then reduced to 5-methyl orsellinic aldehyde by the NRPS-like
CC protein ATEG_03630 (PubMed:24412543). 5-methyl orsellinic aldehyde is
CC then first hydroxylated by the FAD-dependent monooxygenase ATEG_03635
CC and subsequently hydroxylated by the cytochrome P450 monooxygenase
CC ATEG_03631 to produce the unstable polyhydric phenol precursor of
CC azasperpyranones (PubMed:31908094). On the other hand, the polyketide
CC synthase ATEG_07659 is responsible for producing the 3,5-
CC dimethyloctadienone moiety from acetyl-CoA, three malonyl-CoA, and two
CC S-adenosyl methionines (SAM) (Probable). The 3,5-dimethyloctadienone
CC moiety is then loaded onto the SAT domain of ATEG_07661 and extended
CC with four malonyl-CoA and one SAM, which leads to the formation of 2,4-
CC dihydroxy-6-(5,7-dimethyl-2-oxo-trans-3-trans-5-nonadienyl)-3-
CC methylbenzaldehyde (compound 8) after reductive release and aldol
CC condensation (Probable). The FAD-dependent monooxygenase ATEG_07662 is
CC the next enzyme in the biosynthesis sequence and hydroxylates the side
CC chain at the benzylic position of compound 8 (Probable). In Aspergillus
CC nidulans, afoF, the ortholog of the FAD-dependent oxygenase ATEG_07660,
CC is the key enzyme for the biosynthesis of asperfuranone by catalyzing
CC the hydroxylation at C-8 of to prevent the formation of a six-membered
CC ring hemiacetal intermediate and thus facilitatings the formation of a
CC five-membered ring to produce asperfuranone (Probable). In Aspergillus
CC terreus, ATEG_07660 is probably not functional, which leads to the
CC formation of the six-membered ring hemiacetal intermediate
CC presperpyranone instead of asperfuranone (Probable). Finally,
CC ATEG_03636 is involved in the condensation of the polyhydric phenol
CC moiety produced by cluster A and the perasperpyranone precursor
CC produced by cluster B, to yield azasperpyranone A (Probable). Further
CC modifications of azasperpyranone A result in the production of
CC derivatives, including azasperpyranone B to F (PubMed:31908094).
CC {ECO:0000269|PubMed:24412543, ECO:0000269|PubMed:31908094,
CC ECO:0000305|PubMed:31908094}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5-methylorsellinate + ATP + H(+) + NADPH = 2,4-dihydroxy 5,6-
CC dimethylbenzaldehyde + AMP + diphosphate + NADP(+);
CC Xref=Rhea:RHEA:63060, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:146172, ChEBI:CHEBI:146173, ChEBI:CHEBI:456215;
CC Evidence={ECO:0000269|PubMed:24412543, ECO:0000269|PubMed:31908094};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:63061;
CC Evidence={ECO:0000269|PubMed:24412543, ECO:0000269|PubMed:31908094};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:24412543, ECO:0000303|PubMed:31908094}.
CC -!- INDUCTION: Expression is induced by the azasperpyranone cluster A-
CC specific transcription factor ATEG_03638 which is itself regulated by
CC the azasperpyranone transcriptional regulator ATEG_07667.
CC {ECO:0000269|PubMed:31908094}.
CC -!- DOMAIN: Contains three distinct domains: an adenylation (A) domain that
CC activates the substrate amino acid which is subsequently covalently
CC linked as a thioester (aminoacyl-S-PCP) to the 4'-phosphopantetheine
CC prosthetic group of the second domain, the peptidyl carrier protein
CC (PCP) domain, as well as a reductase (R) release domain.
CC {ECO:0000305|PubMed:31908094}.
CC -!- DISRUPTION PHENOTYPE: Abolishes the production of 5-methyl orsellinic
CC aldehyde and azasperpyranone A(AZA-A) but accumulates 5-methyl
CC orsellinic acid. {ECO:0000269|PubMed:31908094}.
CC -!- BIOTECHNOLOGY: Azasperpyranones display potential anti-cancer
CC activities (PubMed:31908094). Azasperpyranones A, C, D, and F exhibit
CC potent growth-inhibitory activity against the A549, HepG2, HCT-116, and
CC HL-60 cell lines, with IC(50) values of 2.39-14.42 mm, respectively
CC (PubMed:31908094). Moreover, azasperpyranone D significantly inhibits
CC HCT-116 xenograft tumor growth in BALB/c-nu mice (PubMed:31908094). In
CC addition, azasperpyranones A and C can bind with four kinds of
CC therapeutic targets for cancer, eEF2K, FGFR, survivin, and TNF-a
CC (PubMed:31908094). {ECO:0000269|PubMed:31908094}.
CC -!- SIMILARITY: Belongs to the adenylate-forming reductase family.
CC {ECO:0000305}.
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DR EMBL; CH476598; EAU35432.1; -; Genomic_DNA.
DR RefSeq; XP_001212808.1; XM_001212808.1.
DR AlphaFoldDB; Q0CRQ4; -.
DR SMR; Q0CRQ4; -.
DR STRING; 33178.CADATEAP00008210; -.
DR EnsemblFungi; EAU35432; EAU35432; ATEG_03630.
DR GeneID; 4318786; -.
DR VEuPathDB; FungiDB:ATEG_03630; -.
DR eggNOG; KOG1176; Eukaryota.
DR eggNOG; KOG1178; Eukaryota.
DR HOGENOM; CLU_002220_2_1_1; -.
DR OMA; YDWKYFR; -.
DR OrthoDB; 229565at2759; -.
DR Proteomes; UP000007963; Unassembled WGS sequence.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR Gene3D; 1.10.1200.10; -; 1.
DR Gene3D; 3.40.50.12780; -; 1.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig.
DR InterPro; IPR042099; ANL_N_sf.
DR InterPro; IPR013120; Far_NAD-bd.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF00501; AMP-binding; 1.
DR Pfam; PF07993; NAD_binding_4; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR PROSITE; PS00455; AMP_BINDING; 1.
PE 1: Evidence at protein level;
KW ATP-binding; NADP; Nucleotide-binding; Oxidoreductase; Phosphopantetheine;
KW Phosphoprotein; Reference proteome.
FT CHAIN 1..1069
FT /note="Adenylate-forming reductase"
FT /id="PRO_0000449882"
FT DOMAIN 576..656
FT /note="Carrier"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 20..391
FT /note="Adenylation (A) domain"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:24412543"
FT REGION 686..1032
FT /note="Reductase (R) domain"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:24412543"
FT BINDING 262
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 357..358
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 362
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 437..440
FT /ligand="AMP"
FT /ligand_id="ChEBI:CHEBI:456215"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 693..696
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 719
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 785..787
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 863
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT BINDING 867
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q6RKB1"
FT MOD_RES 612
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MUTAGEN 612
FT /note="S->A: Abolishes the production of 2,4-dihydroxy 5,6-
FT dimethyl benzaldehyde but accumulates 5-methylorsellinic
FT acid."
FT /evidence="ECO:0000269|PubMed:24412543"
FT MUTAGEN 690
FT /note="T->A: Almost completely abolishes the catalytic
FT activity."
FT /evidence="ECO:0000269|PubMed:24412543"
FT MUTAGEN 863
FT /note="Y->F: Almost completely abolishes the catalytic
FT activity."
FT /evidence="ECO:0000269|PubMed:24412543"
FT MUTAGEN 867
FT /note="K->A: Completely abolishes the catalytic activity."
FT /evidence="ECO:0000269|PubMed:24412543"
SQ SEQUENCE 1069 AA; 118882 MW; 3F7AA98FD7DFA686 CRC64;
MSPIAIDTAP FQRARVNLLH PEDPKAVKSI VQLLQFNAEH NPDHVFCLQL PSKQDDAIGN
PIRITHLQFY RAVSYCTQRL QEEIDGLHGP RVNEDGTVTK CSPVVLFMES NVGLLIHLLA
LMSLGVPVAV LSARLSPTAV QHLMSSIRAQ SVIASPRLKG TIEEAIASDN NTPAIGVRMY
TQRPFEDDLE NSRTLDLPAT NEESHFISEN DRNVLILHSS GTTGLPKPIY QPHRYLLNYS
ECHELGPDDA LGTVLSALPL FHGFGLVAPC LAMTVGKPFM LPPSNTIPTG SLIIELIQSF
QPTALMTVPH ILEEITTLPP EQSISALQPL EFVLCGGGPL KISVAEALAA SGVNLLAHFG
TTETGPLGVV FVPTPDYDWH YWKLRQDINY RLDEVDANSA DGNQYKLTVH PFGWDSAFEI
QDILLSRGAE YKHHLRAVGR KDDLIVLANG EKLVPRVLET LLMQDERVKS AVAFGEGKFE
IGVIVEPTHK VSDEEDFKAA LWAIVLEAGA QMDSHAQVSS PSSIILATPE KPVPRSDKGS
ILRRETYRVY DEEISRVYEV LDRASEETTA LNLQSDSLEE DLKDLIQREI GWKISPSEWL
QDSDLFELGM NSLQAIRLHR LLLSSLPVDS RERVGADFVY RSPSVSKLGA SLRHLAANEN
GHRNDPETEI DELICLNSFI ARQDATVLLT GSTGNLGSNL LAHLTTLPRV KKVICLNRRG
SDTSTAHTDL VERQLAIAKS KGVVIDPESA SKIEVIPCDP SADFFGLPAE VYTHLTAQTT
HILHNAWPMD FKRNVASFQS QFQYLNNLLR VAHDTRLCRP SIKPRFLFVS SIAVVGQYPR
THGTRLIPEV PSDKSSIIED FGYGKAKYVC EEIMRAAADR YPEMQLGIVR VGQMSGSSRT
GYWNPKEHFP TLIKFASMVG QLPAIKQTLS WIAVDNAATV LSDILFAPSL SGIYHLENPI
RQAWQDVLDI FASSLYINTV NVPFDQWLRN VQAAVQELGT EDERMEYDLL AEFLEKDFQR
MATGKVILDT SRSRAVSETL REVGEISEEV VWKYVREWRR AGTLRAPLE
