5EAS_TOBAC
ID 5EAS_TOBAC Reviewed; 548 AA.
AC Q40577;
DT 15-JUL-1998, integrated into UniProtKB/Swiss-Prot.
DT 29-APR-2008, sequence version 3.
DT 03-AUG-2022, entry version 135.
DE RecName: Full=5-epi-aristolochene synthase {ECO:0000303|Ref.3};
DE Short=TEAS {ECO:0000303|Ref.3};
DE EC=4.2.3.61 {ECO:0000269|PubMed:16375847, ECO:0000269|Ref.3};
GN Name=EAS3;
GN and
GN Name=EAS4;
OS Nicotiana tabacum (Common tobacco).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Solanales; Solanaceae; Nicotianoideae; Nicotianeae;
OC Nicotiana.
OX NCBI_TaxID=4097;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA], PROTEIN SEQUENCE OF 56-73, AND
RP INDUCTION.
RC STRAIN=cv. NK 326;
RX PubMed=1438319; DOI=10.1073/pnas.89.22.11088;
RA Facchini P.J., Chappell J.;
RT "Gene family for an elicitor-induced sesquiterpene cyclase in tobacco.";
RL Proc. Natl. Acad. Sci. U.S.A. 89:11088-11092(1992).
RN [2]
RP SEQUENCE REVISION TO 35.
RA O'Maille P.E.;
RL Unpublished observations (APR-2008).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF TYR-520.
RX DOI=10.1021/ja993584h;
RA Rising K.A., Starks C.M., Noel J.P., Chappell J.;
RT "Demonstration of germacrene A as an intermediate in 5-epi-aristolochene
RT synthase catalysis.";
RL J. Am. Chem. Soc. 122:1861-1866(2000).
RN [4]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=16375847; DOI=10.1016/j.abb.2005.10.028;
RA O'Maille P.E., Chappell J., Noel J.P.;
RT "Biosynthetic potential of sesquiterpene synthases: alternative products of
RT tobacco 5-epi-aristolochene synthase.";
RL Arch. Biochem. Biophys. 448:73-82(2006).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) IN COMPLEX WITH MAGNESIUM IONS, AND
RP SEQUENCE REVISION.
RX PubMed=9295271; DOI=10.1126/science.277.5333.1815;
RA Starks C.M., Back K., Chappell J., Noel J.P.;
RT "Structural basis for cyclic terpene biosynthesis by tobacco 5-epi-
RT aristolochene synthase.";
RL Science 277:1815-1820(1997).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH (2E,6E)-FARNESYL
RP DIPHOSPHATE.
RX PubMed=27328867; DOI=10.1038/ja.2016.68;
RA Koo H.J., Vickery C.R., Xu Y., Louie G.V., O'Maille P.E., Bowman M.,
RA Nartey C.M., Burkart M.D., Noel J.P.;
RT "Biosynthetic potential of sesquiterpene synthases: product profiles of
RT Egyptian Henbane premnaspirodiene synthase and related mutants.";
RL J. Antibiot. 69:524-533(2016).
CC -!- FUNCTION: Catalyzes the cyclization of trans,trans-farnesyl diphosphate
CC (FPP) to the bicyclic intermediate 5-epi-aristolochene, initial step in
CC the conversion of FPP to the sesquiterpenoid antifungal phytoalexin
CC capsidiol (Ref.3, PubMed:16375847). Produces germacrene A as an enzyme-
CC bound intermediate that is not released by the enzyme, but is further
CC cyclized to produce the bicyclic 5-epi-aristolochene (Ref.3,
CC PubMed:16375847). {ECO:0000269|PubMed:16375847, ECO:0000269|Ref.3}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E)-farnesyl diphosphate = (+)-5-epi-aristolochene +
CC diphosphate; Xref=Rhea:RHEA:28635, ChEBI:CHEBI:23925,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:175763; EC=4.2.3.61;
CC Evidence={ECO:0000269|PubMed:16375847, ECO:0000269|Ref.3};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:28636;
CC Evidence={ECO:0000269|PubMed:16375847, ECO:0000269|Ref.3};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:9295271};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000269|PubMed:9295271};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.3 uM for 2-trans,6-trans-farnesyl diphosphate
CC {ECO:0000269|Ref.3};
CC -!- PATHWAY: Secondary metabolite biosynthesis; terpenoid biosynthesis.
CC {ECO:0000305}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:9295271}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- INDUCTION: By fungal elicitor. {ECO:0000269|PubMed:1438319}.
CC -!- DOMAIN: The Asp-Asp-Xaa-Xaa-Asp/Glu (DDXXD/E) motif is important for
CC the catalytic activity, presumably through binding to Mg(2+).
CC {ECO:0000305}.
CC -!- MISCELLANEOUS: PubMed:1438319 indicates the presence of at least two
CC genes coding for the same protein.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; L04680; AAA19216.1; -; Unassigned_RNA.
DR PIR; T03714; T03714.
DR PDB; 1HX9; X-ray; 3.50 A; A=1-548.
DR PDB; 1HXA; X-ray; 2.32 A; A=1-548.
DR PDB; 1HXC; X-ray; 2.25 A; A=1-548.
DR PDB; 1HXG; X-ray; 2.90 A; A=1-548.
DR PDB; 3LZ9; X-ray; 2.28 A; A=1-548.
DR PDB; 3M00; X-ray; 2.10 A; A=1-548.
DR PDB; 3M01; X-ray; 2.60 A; A=1-548.
DR PDB; 3M02; X-ray; 2.50 A; A=1-548.
DR PDB; 4DI5; X-ray; 2.30 A; A=14-548.
DR PDB; 4RNQ; X-ray; 2.47 A; A=1-548.
DR PDB; 5DHI; X-ray; 2.25 A; A=13-548.
DR PDB; 5DHK; X-ray; 2.43 A; A=13-548.
DR PDB; 5EAS; X-ray; 2.25 A; A=1-548.
DR PDB; 5EAT; X-ray; 2.80 A; A=1-548.
DR PDB; 5EAU; X-ray; 2.15 A; A=1-548.
DR PDB; 5IK0; X-ray; 2.20 A; A=1-548.
DR PDB; 5IK6; X-ray; 2.30 A; A=1-548.
DR PDB; 5IK9; X-ray; 2.23 A; A=1-548.
DR PDB; 5IKA; X-ray; 2.45 A; A=1-548.
DR PDB; 5IKH; X-ray; 2.10 A; A=1-548.
DR PDB; 5IL3; X-ray; 1.85 A; A=1-548.
DR PDB; 5IL8; X-ray; 2.30 A; A=1-548.
DR PDB; 5ILD; X-ray; 2.12 A; A=1-548.
DR PDB; 5ILH; X-ray; 2.10 A; A=1-548.
DR PDB; 5ILI; X-ray; 1.90 A; A=1-548.
DR PDB; 5ILJ; X-ray; 2.05 A; A=1-548.
DR PDB; 5ILK; X-ray; 2.35 A; A=1-548.
DR PDB; 5ILY; X-ray; 2.45 A; A=1-548.
DR PDB; 5ILZ; X-ray; 1.92 A; A=1-548.
DR PDB; 5IM1; X-ray; 1.88 A; A=1-548.
DR PDBsum; 1HX9; -.
DR PDBsum; 1HXA; -.
DR PDBsum; 1HXC; -.
DR PDBsum; 1HXG; -.
DR PDBsum; 3LZ9; -.
DR PDBsum; 3M00; -.
DR PDBsum; 3M01; -.
DR PDBsum; 3M02; -.
DR PDBsum; 4DI5; -.
DR PDBsum; 4RNQ; -.
DR PDBsum; 5DHI; -.
DR PDBsum; 5DHK; -.
DR PDBsum; 5EAS; -.
DR PDBsum; 5EAT; -.
DR PDBsum; 5EAU; -.
DR PDBsum; 5IK0; -.
DR PDBsum; 5IK6; -.
DR PDBsum; 5IK9; -.
DR PDBsum; 5IKA; -.
DR PDBsum; 5IKH; -.
DR PDBsum; 5IL3; -.
DR PDBsum; 5IL8; -.
DR PDBsum; 5ILD; -.
DR PDBsum; 5ILH; -.
DR PDBsum; 5ILI; -.
DR PDBsum; 5ILJ; -.
DR PDBsum; 5ILK; -.
DR PDBsum; 5ILY; -.
DR PDBsum; 5ILZ; -.
DR PDBsum; 5IM1; -.
DR AlphaFoldDB; Q40577; -.
DR SMR; Q40577; -.
DR KEGG; ag:AAA19216; -.
DR BioCyc; MetaCyc:EAS-MON; -.
DR BRENDA; 4.2.3.61; 3645.
DR BRENDA; 4.2.3.9; 3645.
DR UniPathway; UPA00213; -.
DR EvolutionaryTrace; Q40577; -.
DR Proteomes; UP000084051; Unplaced.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0102698; F:5-epi-aristolochene synthase activity; IDA:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0010333; F:terpene synthase activity; IEA:InterPro.
DR GO; GO:0016102; P:diterpenoid biosynthetic process; IEA:InterPro.
DR GO; GO:0051762; P:sesquiterpene biosynthetic process; IDA:UniProtKB.
DR CDD; cd00684; Terpene_cyclase_plant_C1; 1.
DR Gene3D; 1.10.600.10; -; 1.
DR Gene3D; 1.50.10.130; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR InterPro; IPR044814; Terpene_cyclase_plant_C1.
DR InterPro; IPR001906; Terpene_synth_N.
DR InterPro; IPR036965; Terpene_synth_N_sf.
DR InterPro; IPR005630; Terpene_synthase_metal-bd.
DR InterPro; IPR008930; Terpenoid_cyclase/PrenylTrfase.
DR Pfam; PF01397; Terpene_synth; 1.
DR Pfam; PF03936; Terpene_synth_C; 1.
DR SUPFAM; SSF48239; SSF48239; 1.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Direct protein sequencing; Lyase; Magnesium;
KW Metal-binding; Reference proteome.
FT CHAIN 1..548
FT /note="5-epi-aristolochene synthase"
FT /id="PRO_0000186438"
FT MOTIF 301..305
FT /note="DDXXD motif"
FT /evidence="ECO:0000305"
FT BINDING 264
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /evidence="ECO:0000269|PubMed:27328867"
FT BINDING 301
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /evidence="ECO:0000269|PubMed:27328867"
FT BINDING 301
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:9295271"
FT BINDING 301
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:9295271"
FT BINDING 305
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /evidence="ECO:0000269|PubMed:27328867"
FT BINDING 305
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:9295271"
FT BINDING 305
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:9295271"
FT BINDING 441
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /evidence="ECO:0000269|PubMed:27328867"
FT BINDING 444
FT /ligand="(2E,6E)-farnesyl diphosphate"
FT /ligand_id="ChEBI:CHEBI:175763"
FT /evidence="ECO:0000269|PubMed:27328867"
FT BINDING 444
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:9295271"
FT BINDING 448
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:9295271"
FT BINDING 452
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:9295271"
FT MUTAGEN 520
FT /note="Y->F: Loss of production of aristolochene, and
FT accumulation of the intermediate germacrene A."
FT /evidence="ECO:0000269|Ref.3"
FT CONFLICT 35
FT /note="K -> D (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT CONFLICT 42
FT /note="Y -> YIY (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 44
FT /note="K -> Q (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT CONFLICT 55
FT /note="N -> S (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT CONFLICT 62
FT /note="M -> R (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT CONFLICT 73
FT /note="T -> I (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT CONFLICT 89
FT /note="D -> E (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT CONFLICT 388
FT /note="T -> M (in Ref. 1; AAA19216)"
FT /evidence="ECO:0000305"
FT TURN 26..29
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 36..57
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 64..76
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 80..82
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 84..97
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 104..116
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 123..129
FT /evidence="ECO:0007829|PDB:5IL3"
FT STRAND 134..136
FT /evidence="ECO:0007829|PDB:5EAU"
FT HELIX 138..142
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 144..154
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 162..164
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 167..178
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 179..181
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 186..195
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 203..213
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 215..217
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 223..252
FT /evidence="ECO:0007829|PDB:5IL3"
FT TURN 253..255
FT /evidence="ECO:0007829|PDB:5IK0"
FT HELIX 256..259
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 267..277
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 281..283
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 284..306
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 310..322
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 325..330
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 333..352
FT /evidence="ECO:0007829|PDB:5IL3"
FT TURN 353..357
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 359..361
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 362..385
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 391..398
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 400..402
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 404..412
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 420..427
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 431..454
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 461..469
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 473..494
FT /evidence="ECO:0007829|PDB:5IL3"
FT STRAND 496..498
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 503..505
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 507..519
FT /evidence="ECO:0007829|PDB:5IL3"
FT TURN 520..522
FT /evidence="ECO:0007829|PDB:5IL3"
FT STRAND 523..525
FT /evidence="ECO:0007829|PDB:5IKH"
FT TURN 526..528
FT /evidence="ECO:0007829|PDB:5IL3"
FT TURN 530..533
FT /evidence="ECO:0007829|PDB:5IL3"
FT HELIX 534..542
FT /evidence="ECO:0007829|PDB:5IL3"
SQ SEQUENCE 548 AA; 62987 MW; 9FE25FF361A68BF1 CRC64;
MASAAVANYE EEIVRPVADF SPSLWGDQFL SFSIKNQVAE KYAKEIEALK EQTRNMLLAT
GMKLADTLNL IDTIERLGIS YHFEKEIDDI LDQIYNQNSN CNDLCTSALQ FRLLRQHGFN
ISPEIFSKFQ DENGKFKESL ASDVLGLLNL YEASHVRTHA DDILEDALAF STIHLESAAP
HLKSPLREQV THALEQCLHK GVPRVETRFF ISSIYDKEQS KNNVLLRFAK LDFNLLQMLH
KQELAQVSRW WKDLDFVTTL PYARDRVVEC YFWALGVYFE PQYSQARVML VKTISMISIV
DDTFDAYGTV KELEAYTDAI QRWDINEIDR LPDYMKISYK AILDLYKDYE KELSSAGRSH
IVCHAIERMK EVVRNYNVES TWFIEGYTPP VSEYLSNALA TTTYYYLATT SYLGMKSATE
QDFEWLSKNP KILEASVIIC RVIDDTATYE VEKSRGQIAT GIECCMRDYG ISTKEAMAKF
QNMAETAWKD INEGLLRPTP VSTEFLTPIL NLARIVEVTY IHNLDGYTHP EKVLKPHIIN
LLVDSIKI
