FSA2_FUSSF
ID FSA2_FUSSF Reviewed; 374 AA.
AC A0A0E4AYE7;
DT 30-AUG-2017, integrated into UniProtKB/Swiss-Prot.
DT 24-JUN-2015, sequence version 1.
DT 03-AUG-2022, entry version 9.
DE RecName: Full=Diels-Alderase fsa2 {ECO:0000303|PubMed:28401214};
DE EC=5.5.1.- {ECO:0000269|PubMed:28401214, ECO:0000269|PubMed:34121297};
DE AltName: Full=Fusarisetin A biosynthesis protein 2 {ECO:0000303|PubMed:25770422};
GN Name=fsa2 {ECO:0000303|PubMed:25770422};
OS Fusarium sp. (strain FN080326).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Nectriaceae; Fusarium.
OX NCBI_TaxID=1608308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, DISRUPTION PHENOTYPE, AND
RP PATHWAY.
RX PubMed=25770422; DOI=10.1016/j.bbrc.2015.03.011;
RA Kato N., Nogawa T., Hirota H., Jang J.H., Takahashi S., Ahn J.S., Osada H.;
RT "A new enzyme involved in the control of the stereochemistry in the decalin
RT formation during equisetin biosynthesis.";
RL Biochem. Biophys. Res. Commun. 460:210-215(2015).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX PubMed=28401214; DOI=10.1039/c7cc01929g;
RA Li X., Zheng Q., Yin J., Liu W., Gao S.;
RT "Chemo-enzymatic synthesis of equisetin.";
RL Chem. Commun. (Camb.) 53:4695-4697(2017).
RN [3]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=29972614; DOI=10.1002/anie.201805050;
RA Kato N., Nogawa T., Takita R., Kinugasa K., Kanai M., Uchiyama M.,
RA Osada H., Takahashi S.;
RT "Control of the stereochemical course of [4+2] cycloaddition during trans-
RT decalin formation by Fsa2-family enzymes.";
RL Angew. Chem. Int. Ed. 57:9754-9758(2018).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.17 ANGSTROMS), DOMAIN, FUNCTION, CATALYTIC
RP ACTIVITY, AND MUTAGENESIS OF GLN-80; TRP-216; TRP-332 AND ASN-346.
RX PubMed=34121297; DOI=10.1002/anie.202106186;
RA Fujiyama K., Kato N., Re S., Kinugasa K., Watanabe K., Takita R.,
RA Nogawa T., Hino T., Osada H., Sugita Y., Takahashi S., Nagano S.;
RT "Molecular basis for two stereoselective Diels-Alderases that produce
RT decalin skeletons*.";
RL Angew. Chem. Int. Ed. 60:22401-22410(2021).
CC -!- FUNCTION: Diels-Alderase; part of the gene cluster that mediates the
CC biosynthesis of the HIV-1 integrase inhibitor equisetin and of
CC fusarisetin A, both trans-fused decalin-containing tetramic acids
CC showing also antimicrobial activity (PubMed:25770422). The PKS module
CC of fsa1 together with the enoylreductase fsa3 catalyze the formation of
CC the polyketide unit which is then conjugated to L-serine by the
CC condensation domain of the fsa1 NRPS module (PubMed:25770422). Activity
CC of the Dieckmann cyclase domain (RED) results in release of the
CC Dieckmann product intermediate (PubMed:25770422). Diels-Alderase fsa2
CC is involved in endo-selective Diels-Alder cycloaddition to form the
CC decalin ring, leading to the production of N-desmethylequisetin also
CC called trichosetin (PubMed:25770422, PubMed:28401214, PubMed:29972614,
CC PubMed:34121297). Subsequent N-methylation is carried out by fsa4 to
CC give equisetin (PubMed:25770422). The enzymatic gene responsible for
CC the conversion of equisetin to fusarisetin A has not been identified
CC yet and is probably located outside of the fsa cluster
CC (PubMed:28401214). {ECO:0000269|PubMed:25770422,
CC ECO:0000269|PubMed:28401214, ECO:0000269|PubMed:29972614,
CC ECO:0000269|PubMed:34121297}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(5S)-3-[(2E,6R,8E,10E,12E)-2,6-dimethyltetradeca-2,8,10,12-
CC tetraenoyl]-5-(hydroxymethyl)pyrrolidine-2,4-dione = trichosetin;
CC Xref=Rhea:RHEA:67328, ChEBI:CHEBI:142061, ChEBI:CHEBI:169938;
CC Evidence={ECO:0000269|PubMed:28401214, ECO:0000269|PubMed:34121297};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:67329;
CC Evidence={ECO:0000269|PubMed:28401214, ECO:0000269|PubMed:34121297};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=216 uM for polyenoyltretamic acid {ECO:0000269|PubMed:28401214};
CC pH dependence:
CC Optimum pH is 7. {ECO:0000269|PubMed:28401214};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius.
CC {ECO:0000269|PubMed:28401214};
CC -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000269|PubMed:25770422,
CC ECO:0000269|PubMed:28401214}.
CC -!- DISRUPTION PHENOTYPE: Results in the loss of production of equisetin
CC and fusarisetin A (PubMed:25770422). {ECO:0000269|PubMed:25770422}.
CC -!- SIMILARITY: Belongs to the Diels-Alderase family. {ECO:0000305}.
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DR EMBL; LC025956; BAR40284.1; -; Genomic_DNA.
DR PDB; 7DMN; X-ray; 2.00 A; A=1-374.
DR PDB; 7E22; X-ray; 2.63 A; A=1-374.
DR PDB; 7E5T; X-ray; 2.17 A; A/B/C/D/E/F/G/H=1-374.
DR PDBsum; 7DMN; -.
DR PDBsum; 7E22; -.
DR PDBsum; 7E5T; -.
DR AlphaFoldDB; A0A0E4AYE7; -.
DR SMR; A0A0E4AYE7; -.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
PE 1: Evidence at protein level;
KW 3D-structure; Isomerase.
FT CHAIN 1..374
FT /note="Diels-Alderase fsa2"
FT /id="PRO_0000441292"
FT REGION 1..216
FT /note="Beta-sandwich motif"
FT /evidence="ECO:0000305|PubMed:34121297"
FT REGION 216..374
FT /note="Beta-barrel motif"
FT /evidence="ECO:0000305|PubMed:34121297"
FT MUTAGEN 80
FT /note="Q->A: Slightly decreases enzyme activity."
FT /evidence="ECO:0000269|PubMed:34121297"
FT MUTAGEN 216
FT /note="W->A: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:34121297"
FT MUTAGEN 332
FT /note="W->A: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:34121297"
FT MUTAGEN 346
FT /note="N->A: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:34121297"
FT STRAND 5..10
FT /evidence="ECO:0007829|PDB:7DMN"
FT TURN 11..13
FT /evidence="ECO:0007829|PDB:7E5T"
FT STRAND 14..18
FT /evidence="ECO:0007829|PDB:7E5T"
FT STRAND 45..54
FT /evidence="ECO:0007829|PDB:7DMN"
FT TURN 55..57
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 60..68
FT /evidence="ECO:0007829|PDB:7DMN"
FT HELIX 69..73
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 77..84
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 90..104
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 110..116
FT /evidence="ECO:0007829|PDB:7DMN"
FT TURN 117..120
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 121..127
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 131..140
FT /evidence="ECO:0007829|PDB:7DMN"
FT TURN 141..143
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 144..152
FT /evidence="ECO:0007829|PDB:7DMN"
FT TURN 166..168
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 169..187
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 194..198
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 201..214
FT /evidence="ECO:0007829|PDB:7DMN"
FT HELIX 216..219
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 221..230
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 233..241
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 251..258
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 261..266
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 281..287
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 299..301
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 306..312
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 320..340
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 345..357
FT /evidence="ECO:0007829|PDB:7DMN"
FT STRAND 362..372
FT /evidence="ECO:0007829|PDB:7DMN"
SQ SEQUENCE 374 AA; 40799 MW; 30932F2283A69408 CRC64;
MSNVTVSAFT VDKSISEEHV LPSSFIPGSG NIFPKFTSAI PKTAWELWYF DGISKDDKSS
IVIGVTRNAE GLKHGGFKVQ VFVIWADERT WHRDLFFPES VVSINESGVT DGIWKDATSN
SSISFSCAGD LSKASLVFDV PGVVQGDMHL EALPGDTGLD TDARLGPSVY YVRPIGRASV
KAQLSLYSSD ATAAEQFSLG TSANGGMDRV WSPLSWPQVM TESYYLRTQV GPYAMQIMRI
FPPAGSEDQP STMARLYREG QLVCVAQHVV TREDALMTHD SLILSKQDNS DSEDVVTGGY
RDKNTGYTVE FVEKGNEGQR WKFQVRHERI IWNTPTSRPG PDATGNTGFV EVLCGGTIGE
SYEGVGTGGQ CELS
