NOTF_ASPSM
ID NOTF_ASPSM Reviewed; 452 AA.
AC E0Y3X1;
DT 11-DEC-2019, integrated into UniProtKB/Swiss-Prot.
DT 02-NOV-2010, sequence version 1.
DT 03-AUG-2022, entry version 32.
DE RecName: Full=Deoxybrevianamide E synthase notF {ECO:0000303|PubMed:20722388};
DE EC=2.5.1.109 {ECO:0000269|PubMed:20722388};
DE AltName: Full=Reverse prenyltransferase notF {ECO:0000303|PubMed:20722388};
GN Name=notF {ECO:0000303|PubMed:20722388};
OS Aspergillus sp. (strain MF297-2).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus.
OX NCBI_TaxID=877550;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, PATHWAY, AND
RP MUTAGENESIS OF GLU-108; ARG-122 AND TRP-424.
RC STRAIN=MF297-2;
RX PubMed=20722388; DOI=10.1021/ja1049302;
RA Ding Y., de Wet J.R., Cavalcoli J., Li S., Greshock T.J., Miller K.A.,
RA Finefield J.M., Sunderhaus J.D., McAfoos T.J., Tsukamoto S., Williams R.M.,
RA Sherman D.H.;
RT "Genome-based characterization of two prenylation steps in the assembly of
RT the stephacidin and notoamide anticancer agents in a marine-derived
RT Aspergillus sp.";
RL J. Am. Chem. Soc. 132:12733-12740(2010).
RN [2]
RP BIOTECHNOLOGY.
RX PubMed=17304611; DOI=10.1002/anie.200604381;
RA Kato H., Yoshida T., Tokue T., Nojiri Y., Hirota H., Ohta T.,
RA Williams R.M., Tsukamoto S.;
RT "Notoamides A-D: prenylated indole alkaloids isolated from a marine-derived
RT fungus, Aspergillus sp.";
RL Angew. Chem. Int. Ed. 46:2254-2256(2007).
RN [3]
RP FUNCTION.
RX PubMed=22188465; DOI=10.1021/ja2093212;
RA Li S., Finefield J.M., Sunderhaus J.D., McAfoos T.J., Williams R.M.,
RA Sherman D.H.;
RT "Biochemical characterization of NotB as an FAD-dependent oxidase in the
RT biosynthesis of notoamide indole alkaloids.";
RL J. Am. Chem. Soc. 134:788-791(2012).
RN [4]
RP FUNCTION.
RX PubMed=23213353; DOI=10.1039/c2md20029e;
RA Li S., Anand K., Tran H., Yu F., Finefield J.M., Sunderhaus J.D.,
RA McAfoos T.J., Tsukamoto S., Williams R.M., Sherman D.H.;
RT "Comparative analysis of the biosynthetic systems for fungal
RT bicyclo[2.2.2]diazaoctane indole alkaloids: the (+)/(-)-notoamide,
RT paraherquamide and malbrancheamide pathways.";
RL Med. Chem. Commun. 3:987-996(2012).
CC -!- FUNCTION: Deoxybrevianamide E synthase; part of the gene cluster that
CC mediates the biosynthesis of notoamide, a fungal indole alkaloid that
CC belongs to a family of natural products containing a characteristic
CC bicyclo[2.2.2]diazaoctane core (PubMed:20722388). The first step of
CC notoamide biosynthesis involves coupling of L-proline and L-tryptophan
CC by the bimodular NRPS notE, to produce cyclo-L-tryptophan-L-proline
CC called brevianamide F (PubMed:20722388). The reverse prenyltransferase
CC notF then acts as a deoxybrevianamide E synthase and converts
CC brevianamide F to deoxybrevianamide E via reverse prenylation at C-2 of
CC the indole ring leading to the bicyclo[2.2.2]diazaoctane core
CC (PubMed:20722388). Deoxybrevianamide E is further hydroxylated at C-6
CC of the indole ring, likely catalyzed by the cytochrome P450
CC monooxygenase notG, to yield 6-hydroxy-deoxybrevianamide E (Probable).
CC 6-hydroxy-deoxybrevianamide E is a specific substrate of the
CC prenyltransferase notC for normal prenylation at C-7 to produce 6-
CC hydroxy-7-prenyl-deoxybrevianamide, also called notoamide S
CC (PubMed:20722388). As the proposed pivotal branching point in notoamide
CC biosynthesis, notoamide S can be diverted to notoamide E through an
CC oxidative pyran ring closure putatively catalyzed by either notH
CC cytochrome P450 monooxygenase or the notD FAD-linked oxidoreductase
CC (Probable). This step would be followed by an indole 2,3-epoxidation-
CC initiated pinacol-like rearrangement catalyzed by the notB FAD-
CC dependent monooxygenase leading to the formation of notoamide C and
CC notoamide D (PubMed:22188465). On the other hand notoamide S is
CC converted to notoamide T by notH (or notD), a bifunctional oxidase that
CC also functions as the intramolecular Diels-Alderase responsible for
CC generation of (+)-notoamide T (Probable). To generate antipodal (-)-
CC notoaminide T, notH' (or notD') in Aspergillus versicolor is expected
CC to catalyze a Diels-Alder reaction leading to the opposite
CC stereochemistry (Probable). The remaining oxidoreductase notD (or notH)
CC likely catalyzes the oxidative pyran ring formation to yield (+)-
CC stephacidin A (Probable). The FAD-dependent monooxygenase notI is
CC highly similar to notB and is predicted to catalyze a similar
CC conversion from (+)-stephacidin A to (-)-notoamide B via the 2,3-
CC epoxidation of (+)-stephacidin A followed by a pinacol-type
CC rearrangement (Probable). Finally, it remains unclear which enzyme
CC could be responsible for the final hydroxylation steps leading to
CC notoamide A and sclerotiamide (Probable). {ECO:0000269|PubMed:20722388,
CC ECO:0000269|PubMed:22188465, ECO:0000305|PubMed:23213353}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=brevianamide F + dimethylallyl diphosphate = deoxybrevianamide
CC E + diphosphate; Xref=Rhea:RHEA:35943, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:57623, ChEBI:CHEBI:64530, ChEBI:CHEBI:72948;
CC EC=2.5.1.109; Evidence={ECO:0000269|PubMed:20722388};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:35944;
CC Evidence={ECO:0000269|PubMed:20722388};
CC -!- ACTIVITY REGULATION: Addition of 5 mM Mg(2+), Ca(2+) or Mn(2+) slightly
CC enhances catalysis (about 100-120%) (PubMed:20722388). Significant
CC reduction of enzyme activity (2%-35%) is observed with Cu(2+), Zn(2+),
CC Fe(2+), or Sn(2+) (5 mM) (PubMed:20722388).
CC {ECO:0000269|PubMed:20722388}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=4.33 uM for brevianamide F {ECO:0000269|PubMed:20722388};
CC KM=1.31 uM for dimethylallyl diphosphate (DMAPP)
CC {ECO:0000269|PubMed:20722388};
CC Vmax=0.89 uM/min/mg enzyme toward brevianamide F
CC {ECO:0000269|PubMed:20722388};
CC Vmax=1.18 uM/min/mg enzyme toward dimethylallyl diphosphate (DMAPP)
CC {ECO:0000269|PubMed:20722388};
CC pH dependence:
CC Optimum pH is 6-9. {ECO:0000269|PubMed:20722388};
CC Temperature dependence:
CC Optimum temperature is 4 to 42 degrees Celsius.
CC {ECO:0000269|PubMed:20722388};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000269|PubMed:20722388}.
CC -!- SUBUNIT: Monomer. {ECO:0000250|UniProtKB:I4AY86}.
CC -!- BIOTECHNOLOGY: Notoamides have been shown to exhibit antitumoral
CC activities (PubMed:17304611). Notoamides A-C show moderate cytotoxicity
CC against HeLa and L1210 cells with IC(50) values in the range of 22-52
CC mg/ml, but the IC(50) value of notoamide D is greater than 100 mg/ml
CC (PubMed:17304611). Moreover, notoamide C induces G2/M-cell cycle arrest
CC at a concentration of 6.3 mg/ml (PubMed:17304611).
CC {ECO:0000269|PubMed:17304611}.
CC -!- SIMILARITY: Belongs to the tryptophan dimethylallyltransferase family.
CC {ECO:0000305}.
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DR EMBL; GU564535; ADM34132.1; -; Genomic_DNA.
DR EMBL; HM622670; ADM34139.1; -; Genomic_DNA.
DR PDB; 6VY9; X-ray; 3.19 A; A/B/C/D/E/F/G/H=1-452.
DR PDB; 6VYA; X-ray; 3.00 A; A/B/C/D/E/F/G/H/I/J/K/L/M/N/O/P/Q/R/S/T/U/V/W/X=1-452.
DR PDBsum; 6VY9; -.
DR PDBsum; 6VYA; -.
DR AlphaFoldDB; E0Y3X1; -.
DR SMR; E0Y3X1; -.
DR KEGG; ag:ADM34132; -.
DR BRENDA; 2.5.1.109; 530.
DR GO; GO:0004659; F:prenyltransferase activity; IEA:UniProtKB-KW.
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 3D-structure; Alkaloid metabolism; Prenyltransferase; Transferase.
FT CHAIN 1..452
FT /note="Deoxybrevianamide E synthase notF"
FT /id="PRO_0000448808"
FT REGION 1..38
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 24..38
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 108
FT /ligand="brevianamide F"
FT /ligand_id="ChEBI:CHEBI:64530"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 122
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 212
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 214
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 282
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 284
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 371
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 436
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT BINDING 440
FT /ligand="dimethylallyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:57623"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT SITE 124
FT /note="Required for regioselectivity"
FT /evidence="ECO:0000250|UniProtKB:Q4WAW7"
FT MUTAGEN 108
FT /note="E->D,G: Leads to less than 8% catalytic activity."
FT /evidence="ECO:0000269|PubMed:20722388"
FT MUTAGEN 122
FT /note="R->G,H: Leads to less than 2% catalytic activity."
FT /evidence="ECO:0000269|PubMed:20722388"
FT MUTAGEN 424
FT /note="W->G: Leads to less than 2% catalytic activity."
FT /evidence="ECO:0000269|PubMed:20722388"
FT MUTAGEN 424
FT /note="W->Y: Retains about 25% catalyticactivity."
FT /evidence="ECO:0000269|PubMed:20722388"
FT HELIX 35..41
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 47..66
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 71..84
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 87..89
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 101..103
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 106..113
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 115..117
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 119..125
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 134..137
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 142..153
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 162..169
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 175..181
FT /evidence="ECO:0007829|PDB:6VYA"
FT TURN 186..188
FT /evidence="ECO:0007829|PDB:6VY9"
FT STRAND 197..204
FT /evidence="ECO:0007829|PDB:6VYA"
FT TURN 205..207
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 208..215
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 218..223
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 228..238
FT /evidence="ECO:0007829|PDB:6VYA"
FT TURN 239..242
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 247..260
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 268..274
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 281..289
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 292..299
FT /evidence="ECO:0007829|PDB:6VYA"
FT TURN 300..305
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 309..325
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 353..359
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 361..365
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 367..373
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 379..393
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 396..400
FT /evidence="ECO:0007829|PDB:6VYA"
FT HELIX 402..407
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 411..413
FT /evidence="ECO:0007829|PDB:6VY9"
FT TURN 415..417
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 422..430
FT /evidence="ECO:0007829|PDB:6VYA"
FT TURN 431..433
FT /evidence="ECO:0007829|PDB:6VYA"
FT STRAND 434..441
FT /evidence="ECO:0007829|PDB:6VYA"
SQ SEQUENCE 452 AA; 51608 MW; A2E07E5546E023E4 CRC64;
MTAPELRVDT FRAPEDAPKE PSAQQPRLPS SPSPAQALAS YHHFPTNDQE RWWEETGSLF
SRFLEAGQYG LPQQYQFMFF FMHHLIPALG PYPQKWRSTI SRSGLPIEFS LNFQKGSHRL
LRIGFEPVSF LSGSSQDPFN RIPITDLLNR LSKLQLSNFD TPFFQHLLSK FQLSLSEVRQ
LQKQGSGPDA HPLKSQAAFG FDFNPDGAIL VKGYVFPYLK AKAADVPVGT LIAEAVRTID
VERNQFTHAF GLINDYMQES TGYNEYTFLS CDFVETSEQR LKIYGAHTEV TWAKIAEMWT
LGGRLIEEPE IIAGLARLKQ IWSLLQIGEG SRAFKGGFDY DKSSATDQIA SPIIWNYEIH
PGSRFPVPKF YLPVHGENDL HVARALAQFW DSLGWPEHAC AYPDTLQQLY PDQDISQTTR
LQSWISYSYT AKRGVYMSVY YHSQSTYLWE ED
