NOTE_ASPSM
ID NOTE_ASPSM Reviewed; 2240 AA.
AC E1ACQ0;
DT 11-DEC-2019, integrated into UniProtKB/Swiss-Prot.
DT 02-NOV-2010, sequence version 1.
DT 25-MAY-2022, entry version 48.
DE RecName: Full=Nonribisomal peptide synthase notE {ECO:0000303|PubMed:20722388};
DE Short=NRPS notE {ECO:0000303|PubMed:20722388};
DE EC=6.3.1.- {ECO:0000305|PubMed:20722388};
DE AltName: Full=Notoamide biosynthesis cluster protein E {ECO:0000303|PubMed:20722388};
GN Name=notE {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, DOMAIN, AND PATHWAY.
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: Nonribisomal peptide 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=2 ATP + L-proline + L-tryptophan = 2 AMP + brevianamide F + 2
CC diphosphate + 2 H(+); Xref=Rhea:RHEA:35935, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, ChEBI:CHEBI:57912,
CC ChEBI:CHEBI:60039, ChEBI:CHEBI:64530, ChEBI:CHEBI:456215;
CC Evidence={ECO:0000305|PubMed:20722388};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:35936;
CC Evidence={ECO:0000305|PubMed:20722388};
CC -!- PATHWAY: Alkaloid biosynthesis. {ECO:0000305|PubMed:20722388}.
CC -!- DOMAIN: NRP synthetases are composed of discrete domains (adenylation
CC (A), thiolation (T) or peptidyl carrier protein (PCP) and condensation
CC (C) domains) which when grouped together are referred to as a single
CC module. Each module is responsible for the recognition (via the A
CC domain) and incorporation of a single amino acid into the growing
CC peptide product. Thus, an NRP synthetase is generally composed of one
CC or more modules and can terminate in a thioesterase domain (TE) that
CC releases the newly synthesized peptide from the enzyme. Occasionally,
CC epimerase (E) domains (responsible for L- to D-amino acid conversion)
CC are present within the NRP synthetase (By similarity). NotE as the
CC following architecture: A1-T1-C1-A2-T2-C2. The presence of two intact
CC modules suggests that the two modules condense L-tryptophan and L-
CC phenylalanine together. The C-terminal condensation domain might be
CC responsible for cyclization of the dipeptide to form the
CC diketopiperazine structure (Probable). {ECO:0000250|UniProtKB:Q4WMJ7,
CC ECO:0000305|PubMed:20722388}.
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 NRP synthetase family. {ECO:0000305}.
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DR EMBL; HM622670; ADM34138.1; -; Genomic_DNA.
DR AlphaFoldDB; E1ACQ0; -.
DR SMR; E1ACQ0; -.
DR GO; GO:0016874; F:ligase activity; IEA:UniProtKB-KW.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 1.10.1200.10; -; 2.
DR Gene3D; 3.30.300.30; -; 2.
DR Gene3D; 3.30.559.10; -; 2.
DR Gene3D; 3.40.50.12780; -; 2.
DR InterPro; IPR010071; AA_adenyl_domain.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR045851; AMP-bd_C_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig.
DR InterPro; IPR042099; ANL_N_sf.
DR InterPro; IPR023213; CAT-like_dom_sf.
DR InterPro; IPR001242; Condensatn.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR006162; Ppantetheine_attach_site.
DR Pfam; PF00501; AMP-binding; 2.
DR Pfam; PF00668; Condensation; 2.
DR Pfam; PF00550; PP-binding; 2.
DR SUPFAM; SSF47336; SSF47336; 2.
DR TIGRFAMs; TIGR01733; AA-adenyl-dom; 2.
DR PROSITE; PS00455; AMP_BINDING; 2.
DR PROSITE; PS50075; CARRIER; 2.
DR PROSITE; PS00012; PHOSPHOPANTETHEINE; 1.
PE 1: Evidence at protein level;
KW Alkaloid metabolism; Ligase; Phosphopantetheine; Phosphoprotein; Repeat.
FT CHAIN 1..2240
FT /note="Nonribisomal peptide synthase notE"
FT /id="PRO_0000448811"
FT DOMAIN 616..692
FT /note="Carrier 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258,
FT ECO:0000305|PubMed:20722388"
FT DOMAIN 1700..1776
FT /note="Carrier 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258,
FT ECO:0000305|PubMed:20722388"
FT REGION 22..52
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 85..484
FT /note="Adenylation 1"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:20722388"
FT REGION 732..1144
FT /note="Condensation 1"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:20722388"
FT REGION 1167..1564
FT /note="Adenylation 2"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:20722388"
FT REGION 1845..2159
FT /note="Condensation 2"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:20722388"
FT REGION 2008..2027
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 22..49
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT MOD_RES 653
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MOD_RES 1737
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
SQ SEQUENCE 2240 AA; 245924 MW; A9AEE520D4C774AB CRC64;
MDSIKLTESH QGLSVLHAKQ ATETMRETLS SSSSPLSLSS ITSPLSSASE PPALGEIQAR
VSESTLFSNA QVPEFWETCV HEVIQQRCRE APESSAVAAW DGSFTYSELD ELSNRLASSL
ILLGVKAETF VPICMEKSRW ATVAILGVMK AGGAFTLLDP SYPLPRLKTI CEELSSLVVL
SSTTQSERCT QLANVIVVEH LCRAWHPGAY LAQSPATVCP SNVLYVAFTS GSTGKPKGVL
IEHRAYSTGA REHLRVFQID QTSRVLQFSS YAFDVNIMET LSTLMAGGCL CVVGEAQRSD
VSLFAEAVDE LQVSHAMLTP SFARTVPWEN VRHLRVLILG GEEMRESDAA ICAERGVRLI
NAYGAAECSV NATARPGVQP GDNLSTIGHG TGAIAWIIDP DDPERQMGPG TAGELLLEGP
IVGRGYLNSP DMTDRVFIDP PTWLRQLRMM DYQHQLYRTG DLAVQDSTGA LTLLGRKEGQ
VKIRGQRVEV AEIEQHIDQV LTAATEVVVE KVTPECDQRD VLMAFVQTGR AAQGWTEGSP
FFLPPRPASI QEFSAAQSQL REQLPSYMIP AIFIGVSCIP RTPSGKADRR LLRMTAARLS
REELQAFAGS PVHSRPPTTA TEHALQQLYA DVLELPIMRI SMEDSFVRLG GDSIMAVRLV
GAARQAGLVL DIRDVLGTAR LEEQARKATP VSEETPCEAY VPFSMLGSRC TDRDEVLRVA
AEQCGTSPSE IEDIFPCTPL QEGMLALSSS QPQMYVGQIV FGMPEDVDVS QFKAAWQSTA
DATPILRTRI IHTPQGLLQV VLRGKLAWEN YNEHPEACAA DVGSQIGSPG APLIRFALGD
GDHRGEFTLT VHHAVWDAWT MRLIHDALER SFQGEMTKKH PFHPFIQYLQ QVDGATMDDF
WRTELADLEA PSFPALPSTQ HRPSPTAMLR HTVEKIEVVP RIHTMASYIH LAWSLLVAHY
TDSTEAVYGA IVSGRNSPVA AINELAGPTI ATVPVRVRVS PEDTVSAALE QIQTCMVRMV
PHEQAGLLRI AKASPDAARA CSFQSHLNIQ VVEQEHRLLP ARRGIASTGM ELTRFSSYAL
NLMLQLSPDN TSVTVDIAYD PQVLSAWEVD RMIHQWEHIL RQICREPSGS LQELDFASPQ
DRDLLRLRNS ETPTVDWRCL HDLVLAQEAR QPSREAVSAW DGDFTYRELA ELSSNFARLL
NLFAVGRGSF VPICMDKSRW AIVSILAVLQ AGATCVLLDP QHPRQRMQDT IAGLSVPVLV
NAPSTAPVTK GLCAIELCVS AKLTEQLWTN AYGSRFQTHV DPDDLAFLIF TSGSTGVPKG
IAMPHCTVSS SIYHNSAPMM FDADTRALHF SSYAFDVSIY EIFTTLASGG CVCVPSEFQR
TNELADFIQQ RAVNWTFLTP STAQSLHPSE VPGVATLVLG GEAVTPDHVK TWAPGRSLIN
GYGPAEATIC AVGPMPEHGW DPGNIGHVVG GVGWVTIPSD PSRLAAIGAI GELLLEGPFL
ARGYLNQHEA TAASFISPPP WHRTLLPDCD AETTRLYRTG DLVQYEEDGS IRYIGRRDTQ
LKLRGQRIDL GEIETQLRRS FPGVHDVVAE AIQLPILQDR AALVAFIGCQ EAQVTESAVG
EQVLSAVDES FQHTVSLAQT RLQAILPPYM LPSVFFPLAH CPKTLTGKTD RRYLRQAVLA
LPPHELQRYR VAGRQKARIP VSRGPELRLQ SIWADLLRIP CDDIGSDDTF LLHGGDSVAA
MRMVALARRA DFTFRVTDVL SNCTLSELAR CTGEEPCLTD GDGTLPTTHE FESGHKMVDS
PVSADYHTGM IGTELEMEND AIAVYPTTQA QSFLIKRYPW THWRFSFHGE VSVERLRTAC
ARLVAAHSIM RTLFVAGAGG ERVRHVVMKE LDIPLHTGTT HKNLVDYCQS ICDAEQEMDV
LEAVLPTRLT LISDALQTSH IFILRLSHAQ YDGICVPKIF ADLEALYNGT EPIAPTRFER
YLDERRWYSG ERARAFWKEY LAGSSPPCTM PVKATPPTDS DDSRPSAARS VISASQTVRC
TAIPFQVTLA TVVKAAACLV LARLTGRTDI TVGQTVNGRS LPQPWVSEVV GPCVNYIPFR
ATLSESMSIQ DYLVHMQSQH NRCIHYDGAE LDTIIKNCTT WDPSTEFGFI LQHQNIDMDL
SLTLDGNRCA SCASSGRLRP SNEVWICSTP SPSGVDLDVV ASSQTLTADA AKNLVDDIAD
MIQTLLYNLE TPLRDVVEFD
