TRI5_FUSSP
ID TRI5_FUSSP Reviewed; 374 AA.
AC P13513; Q7LP67;
DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1990, sequence version 1.
DT 03-AUG-2022, entry version 100.
DE RecName: Full=Trichodiene synthase {ECO:0000303|PubMed:3800398};
DE EC=4.2.3.6 {ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17678871, ECO:0000269|PubMed:3800398, ECO:0000269|PubMed:7873527, ECO:0000269|PubMed:8823172};
DE AltName: Full=Core trichothecene cluster (CTC) protein 5 {ECO:0000303|PubMed:8593041};
DE AltName: Full=Sesquiterpene cyclase TRI5 {ECO:0000303|PubMed:3800398};
DE Short=TS {ECO:0000303|PubMed:3800398};
GN Name=TRI5 {ECO:0000303|PubMed:3800398}; Synonyms=TOX 5;
OS Fusarium sporotrichioides.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Nectriaceae; Fusarium.
OX NCBI_TaxID=5514;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PARTIAL PROTEIN SEQUENCE.
RC STRAIN=ATCC 24631 / NRRL 3299;
RX PubMed=2777086; DOI=10.1016/0378-1119(89)90098-x;
RA Hohn T.M., Beremand P.D.;
RT "Isolation and nucleotide sequence of a sesquiterpene cyclase gene from the
RT trichothecene-producing fungus Fusarium sporotrichioides.";
RL Gene 79:131-138(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=ATCC 24631 / NRRL 3299;
RX PubMed=11352533; DOI=10.1006/fgbi.2001.1256;
RA Brown D.W., McCormick S.P., Alexander N.J., Proctor R.H., Desjardins A.E.;
RT "A genetic and biochemical approach to study trichothecene diversity in
RT Fusarium sporotrichioides and Fusarium graminearum.";
RL Fungal Genet. Biol. 32:121-133(2001).
RN [3]
RP NUCLEOTIDE SEQUENCE [MRNA].
RA Ren Q., Tag A., Peplow A., Lai H., Kupfer D., Peterson A., Beremand M.,
RA Roe B.;
RT "Fusarium sporotrichioides trichodiene synthase (TRI5) gene cDNA.";
RL Submitted (APR-2001) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND
RP PATHWAY.
RX PubMed=3800398; DOI=10.1016/0003-9861(86)90386-3;
RA Hohn T.M., Vanmiddlesworth F.;
RT "Purification and characterization of the sesquiterpene cyclase trichodiene
RT synthetase from Fusarium sporotrichioides.";
RL Arch. Biochem. Biophys. 251:756-761(1986).
RN [5]
RP FUNCTION.
RX PubMed=2817906; DOI=10.1016/0003-9861(89)90353-6;
RA Hohn T.M., Plattner R.D.;
RT "Expression of the trichodiene synthase gene of Fusarium sporotrichioides
RT in Escherichia coli results in sesquiterpene production.";
RL Arch. Biochem. Biophys. 275:92-97(1989).
RN [6]
RP INDUCTION.
RX PubMed=16347944; DOI=10.1128/aem.55.6.1500-1503.1989;
RA Hohn T.M., Beremand M.N.;
RT "Regulation of trichodiene synthase in Fusarium sporotrichioides and
RT Gibberella pulicaris (Fusarium sambucinum).";
RL Appl. Environ. Microbiol. 55:1500-1503(1989).
RN [7]
RP FUNCTION.
RX PubMed=2317042; DOI=10.1128/aem.56.3.702-706.1990;
RA McCormick S.P., Taylor S.L., Plattner R.D., Beremand M.N.;
RT "Bioconversion of possible T-2 toxin precursors by a mutant strain of
RT Fusarium sporotrichioides NRRL 3299.";
RL Appl. Environ. Microbiol. 56:702-706(1990).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF CYS-146; CYS-190; ARG-304
RP AND TYR-305.
RX PubMed=7873527; DOI=10.1021/bi00008a011;
RA Cane D.E., Shim J.H., Xue Q., Fitzsimons B.C., Hohn T.M.;
RT "Trichodiene synthase. Identification of active site residues by site-
RT directed mutagenesis.";
RL Biochemistry 34:2480-2488(1995).
RN [9]
RP FUNCTION.
RX PubMed=7651333; DOI=10.1007/bf02456618;
RA Hohn T.M., Desjardins A.E., McCormick S.P.;
RT "The Tri4 gene of Fusarium sporotrichioides encodes a cytochrome P450
RT monooxygenase involved in trichothecene biosynthesis.";
RL Mol. Gen. Genet. 248:95-102(1995).
RN [10]
RP FUNCTION.
RX PubMed=8593041; DOI=10.1128/aem.62.2.353-359.1996;
RA McCormick S.P., Hohn T.M., Desjardins A.E.;
RT "Isolation and characterization of Tri3, a gene encoding 15-O-
RT acetyltransferase from Fusarium sporotrichioides.";
RL Appl. Environ. Microbiol. 62:353-359(1996).
RN [11]
RP FUNCTION, DOMAIN, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, CATALYTIC
RP ACTIVITY, AND MUTAGENESIS OF ASP-100; ASP-101 AND ASP-104.
RX PubMed=8823172; DOI=10.1021/bi961344y;
RA Cane D.E., Xue Q., Fitzsimons B.C.;
RT "Trichodiene synthase. Probing the role of the highly conserved aspartate-
RT rich region by site-directed mutagenesis.";
RL Biochemistry 35:12369-12376(1996).
RN [12]
RP FUNCTION.
RX PubMed=9435078; DOI=10.1128/aem.64.1.221-225.1998;
RA Alexander N.J., Hohn T.M., McCormick S.P.;
RT "The TRI11 gene of Fusarium sporotrichioides encodes a cytochrome P-450
RT monooxygenase required for C-15 hydroxylation in trichothecene
RT biosynthesis.";
RL Appl. Environ. Microbiol. 64:221-225(1998).
RN [13]
RP FUNCTION.
RX PubMed=10583973; DOI=10.1128/aem.65.12.5252-5256.1999;
RA McCormick S.P., Alexander N.J., Trapp S.E., Hohn T.M.;
RT "Disruption of TRI101, the gene encoding trichothecene 3-O-
RT acetyltransferase, from Fusarium sporotrichioides.";
RL Appl. Environ. Microbiol. 65:5252-5256(1999).
RN [14]
RP FUNCTION.
RX PubMed=12039755; DOI=10.1128/aem.68.6.2959-2964.2002;
RA McCormick S.P., Alexander N.J.;
RT "Fusarium Tri8 encodes a trichothecene C-3 esterase.";
RL Appl. Environ. Microbiol. 68:2959-2964(2002).
RN [15]
RP FUNCTION.
RX PubMed=12135578; DOI=10.1016/s1087-1845(02)00021-x;
RA Brown D.W., McCormick S.P., Alexander N.J., Proctor R.H., Desjardins A.E.;
RT "Inactivation of a cytochrome P-450 is a determinant of trichothecene
RT diversity in Fusarium species.";
RL Fungal Genet. Biol. 36:224-233(2002).
RN [16]
RP FUNCTION.
RX PubMed=12620849; DOI=10.1128/aem.69.3.1607-1613.2003;
RA Meek I.B., Peplow A.W., Ake C. Jr., Phillips T.D., Beremand M.N.;
RT "Tri1 encodes the cytochrome P450 monooxygenase for C-8 hydroxylation
RT during trichothecene biosynthesis in Fusarium sporotrichioides and resides
RT upstream of another new Tri gene.";
RL Appl. Environ. Microbiol. 69:1607-1613(2003).
RN [17]
RP INDUCTION.
RX PubMed=12732543; DOI=10.1128/aem.69.5.2731-2736.2003;
RA Peplow A.W., Tag A.G., Garifullina G.F., Beremand M.N.;
RT "Identification of new genes positively regulated by Tri10 and a regulatory
RT network for trichothecene mycotoxin production.";
RL Appl. Environ. Microbiol. 69:2731-2736(2003).
RN [18]
RP FUNCTION.
RX PubMed=14532047; DOI=10.1128/aem.69.10.5935-5940.2003;
RA Peplow A.W., Meek I.B., Wiles M.C., Phillips T.D., Beremand M.N.;
RT "Tri16 is required for esterification of position C-8 during trichothecene
RT mycotoxin production by Fusarium sporotrichioides.";
RL Appl. Environ. Microbiol. 69:5935-5940(2003).
RN [19]
RP FUNCTION.
RX PubMed=16917519; DOI=10.1139/w06-011;
RA McCormick S.P., Alexander N.J., Proctor R.H.;
RT "Fusarium Tri4 encodes a multifunctional oxygenase required for
RT trichothecene biosynthesis.";
RL Can. J. Microbiol. 52:636-642(2006).
RN [20] {ECO:0007744|PDB:1JFA, ECO:0007744|PDB:1JFG}
RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS).
RX PubMed=11698643; DOI=10.1073/pnas.231313098;
RA Rynkiewicz M.J., Cane D.E., Christianson D.W.;
RT "Structure of trichodiene synthase from Fusarium sporotrichioides provides
RT mechanistic inferences on the terpene cyclization cascade.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:13543-13548(2001).
RN [21]
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS), AND MUTAGENESIS OF ASP-100.
RX PubMed=11827517; DOI=10.1021/bi011960g;
RA Rynkiewicz M.J., Cane D.E., Christianson D.W.;
RT "X-ray crystal structures of D100E trichodiene synthase and its
RT pyrophosphate complex reveal the basis for terpene product diversity.";
RL Biochemistry 41:1732-1741(2002).
RN [22] {ECO:0007744|PDB:1KIY, ECO:0007744|PDB:1KIZ}
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) IN COMPLEX WITH MAGNESIUM, COFACTOR,
RP MUTAGENESIS OF ARG-304, AND CATALYTIC ACTIVITY.
RX PubMed=16171386; DOI=10.1021/bi0510476;
RA Vedula L.S., Cane D.E., Christianson D.W.;
RT "Role of arginine-304 in the diphosphate-triggered active site closure
RT mechanism of trichodiene synthase.";
RL Biochemistry 44:12719-12727(2005).
RN [23] {ECO:0007744|PDB:1YJ4, ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR, ECO:0007744|PDB:1YYS, ECO:0007744|PDB:1YYT, ECO:0007744|PDB:1YYU}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH MAGNESIUM AND
RP INHIBITOR, COFACTOR, AND MUTAGENESIS OF TYR-305.
RX PubMed=15835903; DOI=10.1021/bi050059o;
RA Vedula L.S., Rynkiewicz M.J., Pyun H.J., Coates R.M., Cane D.E.,
RA Christianson D.W.;
RT "Molecular recognition of the substrate diphosphate group governs product
RT diversity in trichodiene synthase mutants.";
RL Biochemistry 44:6153-6163(2005).
RN [24] {ECO:0007744|PDB:2Q9Y, ECO:0007744|PDB:2Q9Z}
RP X-RAY CRYSTALLOGRAPHY (2.85 ANGSTROMS) IN COMPLEX WITH MAGNESIUM AND
RP INHIBITOR, CATALYTIC ACTIVITY, AND ACTIVITY REGULATION.
RX PubMed=17678871; DOI=10.1016/j.abb.2007.06.016;
RA Vedula L.S., Zhao Y., Coates R.M., Koyama T., Cane D.E., Christianson D.W.;
RT "Exploring biosynthetic diversity with trichodiene synthase.";
RL Arch. Biochem. Biophys. 466:260-266(2007).
RN [25] {ECO:0007744|PDB:2PS4, ECO:0007744|PDB:2PS5, ECO:0007744|PDB:2PS6, ECO:0007744|PDB:2PS7, ECO:0007744|PDB:2PS8}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH MAGNESIUM, AND
RP MUTAGENESIS OF ASN-225; SER-229 AND TYR-295.
RX PubMed=17996718; DOI=10.1016/j.abb.2007.10.015;
RA Vedula L.S., Jiang J., Zakharian T., Cane D.E., Christianson D.W.;
RT "Structural and mechanistic analysis of trichodiene synthase using site-
RT directed mutagenesis: probing the catalytic function of tyrosine-295 and
RT the asparagine-225/serine-229/glutamate-233-Mg2+B motif.";
RL Arch. Biochem. Biophys. 469:184-194(2008).
CC -!- FUNCTION: Trichodiene synthase; part of the core gene cluster that
CC mediates the biosynthesis of trichothecenes, a very large family of
CC chemically related bicyclic sesquiterpene compounds acting as
CC mycotoxins, including T2-toxin (PubMed:11352533, PubMed:2817906). The
CC biosynthesis of trichothecenes begins with the cyclization of farnesyl
CC diphosphate to trichodiene and is catalyzed by the trichodiene synthase
CC TRI5 (PubMed:3800398, PubMed:7873527, PubMed:8823172). Trichodiene
CC undergoes a series of oxygenations catalyzed by the cytochrome P450
CC monooxygenase TRI4 (PubMed:7651333). TRI4 controls the addition of four
CC oxygens at C-2, C-3, C-11, and the C-12, C-13-epoxide to form the
CC intermediate isotrichotriol (PubMed:16917519). Isotrichotriol then
CC undergoes a non-enzymatic isomerization and cyclization to form
CC isotrichodermol (PubMed:2317042). During this process, the oxygen at
CC the C-2 position becomes the pyran ring oxygen and the hydroxyl group
CC at C-11 is lost (PubMed:2317042). More complex type A trichothecenes
CC are built by modifying isotrichodermol through a series of paired
CC hydroxylation and acetylation or acylation steps (PubMed:11352533).
CC Isotrichodermol is converted to isotrichodermin by the
CC acetyltransferase TRI101 (PubMed:10583973). TRI101 encodes a C-3
CC transacetylase that acts as a self-protection or resistance factor
CC during biosynthesis and that the presence of a free C-3 hydroxyl group
CC is a key component of Fusarium trichothecene phytotoxicity
CC (PubMed:10583973). A second hydroxyl group is added to C-15 by the
CC trichothecene C-15 hydroxylase TRI11, producing 15-decalonectrin, which
CC is then acetylated by TRI3, producing calonectrin (PubMed:9435078,
CC PubMed:8593041). A third hydroxyl group is added at C-4 by the
CC cytochrome P450 monooxygenase TRI13, converting calonectrin to 3,15-
CC diacetoxyspirpenol, which is subsequently acetylated by the
CC acetyltransferase TRI7 (PubMed:12135578, PubMed:11352533). A fourth
CC hydroxyl group is added to C-8 by the cytochrome P450 monooxygenase
CC TRI1, followed by the addition of an isovaleryl moiety by TRI16
CC (PubMed:12620849, PubMed:14532047). Finally, the acetyl group is
CC removed from the C-3 position by the trichothecene C-3 esterase TRI8 to
CC produce T-2 toxin (PubMed:12039755). {ECO:0000269|PubMed:10583973,
CC ECO:0000269|PubMed:11352533, ECO:0000269|PubMed:12039755,
CC ECO:0000269|PubMed:12135578, ECO:0000269|PubMed:12620849,
CC ECO:0000269|PubMed:14532047, ECO:0000269|PubMed:16917519,
CC ECO:0000269|PubMed:2317042, ECO:0000269|PubMed:3800398,
CC ECO:0000269|PubMed:7651333, ECO:0000269|PubMed:8593041,
CC ECO:0000269|PubMed:9435078}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E)-farnesyl diphosphate = diphosphate + trichodiene;
CC Xref=Rhea:RHEA:12052, ChEBI:CHEBI:15861, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:175763; EC=4.2.3.6;
CC Evidence={ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17678871,
CC ECO:0000269|PubMed:3800398, ECO:0000269|PubMed:7873527,
CC ECO:0000269|PubMed:8823172};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:15835903, ECO:0000269|PubMed:16171386,
CC ECO:0000269|PubMed:17678871, ECO:0000269|PubMed:17996718,
CC ECO:0000269|PubMed:3800398, ECO:0000269|PubMed:8823172};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:8823172};
CC Note=Some of the cofactor binding sites show unusual localization
CC within the protein. {ECO:0000305};
CC -!- ACTIVITY REGULATION: Benzyl triethylammonium cation (BTAC) acts as a
CC competitive inhibitor of trichodiene synthase reaction in the presence
CC of pyrophosphate (PPi) (PubMed:17678871).
CC {ECO:0000269|PubMed:17678871}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=62.0 nM for farnesyl pyrophosphate {ECO:0000269|PubMed:3800398,
CC ECO:0000269|PubMed:7873527};
CC KM=78.0 nM for Mg(2+) {ECO:0000269|PubMed:8823172};
CC KM=84.8 nM for Mn(2+) {ECO:0000269|PubMed:8823172};
CC pH dependence:
CC Optimum pH is 6.75-7.75. {ECO:0000269|PubMed:3800398};
CC -!- PATHWAY: Sesquiterpene biosynthesis; trichothecene biosynthesis.
CC {ECO:0000269|PubMed:3800398}.
CC -!- INDUCTION: Expression is positively regulated by the trichothecene
CC cluster-specific transcription activator TRI10 (PubMed:12732543).
CC Expression is induced between 18h and 21h growth on GYEP medium
CC (PubMed:16347944). The initial detection of trichothecenes occurs
CC several hours after the initial detection of TRI5 (PubMed:16347944).
CC The initiation of trichothecene biosynthesis occurs with a high
CC concentration of glucose remaining in the culture medium
CC (PubMed:16347944). {ECO:0000269|PubMed:12732543,
CC ECO:0000269|PubMed:16347944}.
CC -!- MISCELLANEOUS: Trichothecenes are sesquiterpenoid toxins that act by
CC inhibiting protein biosynthesis. {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the trichodiene synthase family. {ECO:0000305}.
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DR EMBL; AF364179; AAD13657.1; -; Genomic_DNA.
DR EMBL; AF359360; AAK33074.1; -; Genomic_DNA.
DR EMBL; AY032745; AAK77935.1; -; mRNA.
DR PIR; JU0064; SYFUTP.
DR PDB; 1JFA; X-ray; 2.50 A; A/B=1-374.
DR PDB; 1JFG; X-ray; 2.50 A; A/B=1-374.
DR PDB; 1KIY; X-ray; 2.40 A; A/B=1-374.
DR PDB; 1KIZ; X-ray; 2.60 A; A/B=1-374.
DR PDB; 1YJ4; X-ray; 2.30 A; A/B=1-374.
DR PDB; 1YYQ; X-ray; 2.10 A; A/B=1-374.
DR PDB; 1YYR; X-ray; 2.50 A; A/B=1-374.
DR PDB; 1YYS; X-ray; 2.75 A; A/B=1-374.
DR PDB; 1YYT; X-ray; 2.90 A; A/B=1-374.
DR PDB; 1YYU; X-ray; 2.95 A; A/B=1-374.
DR PDB; 2AEK; X-ray; 2.90 A; A/B=1-374.
DR PDB; 2AEL; X-ray; 2.50 A; A/B=1-374.
DR PDB; 2AET; X-ray; 2.75 A; A/B=1-374.
DR PDB; 2PS4; X-ray; 2.46 A; A/B=1-374.
DR PDB; 2PS5; X-ray; 2.10 A; A/B=1-374.
DR PDB; 2PS6; X-ray; 2.60 A; A/B=1-374.
DR PDB; 2PS7; X-ray; 2.35 A; A/B=1-374.
DR PDB; 2PS8; X-ray; 2.67 A; A/B=1-374.
DR PDB; 2Q9Y; X-ray; 2.85 A; A/B=1-374.
DR PDB; 2Q9Z; X-ray; 2.95 A; A/B=1-374.
DR PDBsum; 1JFA; -.
DR PDBsum; 1JFG; -.
DR PDBsum; 1KIY; -.
DR PDBsum; 1KIZ; -.
DR PDBsum; 1YJ4; -.
DR PDBsum; 1YYQ; -.
DR PDBsum; 1YYR; -.
DR PDBsum; 1YYS; -.
DR PDBsum; 1YYT; -.
DR PDBsum; 1YYU; -.
DR PDBsum; 2AEK; -.
DR PDBsum; 2AEL; -.
DR PDBsum; 2AET; -.
DR PDBsum; 2PS4; -.
DR PDBsum; 2PS5; -.
DR PDBsum; 2PS6; -.
DR PDBsum; 2PS7; -.
DR PDBsum; 2PS8; -.
DR PDBsum; 2Q9Y; -.
DR PDBsum; 2Q9Z; -.
DR AlphaFoldDB; P13513; -.
DR SMR; P13513; -.
DR KEGG; ag:AAD13657; -.
DR BRENDA; 4.2.3.6; 2364.
DR UniPathway; UPA00267; -.
DR EvolutionaryTrace; P13513; -.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0045482; F:trichodiene synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0016106; P:sesquiterpenoid biosynthetic process; IEA:InterPro.
DR Gene3D; 1.10.600.10; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR InterPro; IPR010458; TRI5_ascomyc.
DR InterPro; IPR024652; Trichodiene_synth.
DR Pfam; PF06330; TRI5; 1.
DR PIRSF; PIRSF001388; TRI5; 1.
DR SFLD; SFLDG01021; Trichodiene_Synthase_Like; 1.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Lyase; Magnesium; Metal-binding.
FT CHAIN 1..374
FT /note="Trichodiene synthase"
FT /id="PRO_0000221584"
FT REGION 100..104
FT /note="Aspartate-rich domain"
FT /evidence="ECO:0000303|PubMed:8823172"
FT BINDING 100
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:17996718,
FT ECO:0007744|PDB:2PS7"
FT BINDING 164
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:17996718,
FT ECO:0007744|PDB:2PS7"
FT BINDING 225
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:15835903,
FT ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718,
FT ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR,
FT ECO:0007744|PDB:1YYU, ECO:0007744|PDB:2AET,
FT ECO:0007744|PDB:2PS8"
FT BINDING 229
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:15835903,
FT ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718,
FT ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR,
FT ECO:0007744|PDB:1YYU, ECO:0007744|PDB:2AET,
FT ECO:0007744|PDB:2PS8"
FT BINDING 233
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:15835903,
FT ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718,
FT ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR,
FT ECO:0007744|PDB:1YYU, ECO:0007744|PDB:2AET,
FT ECO:0007744|PDB:2PS8"
FT BINDING 239
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:16171386,
FT ECO:0000269|PubMed:17996718, ECO:0007744|PDB:2AEK,
FT ECO:0007744|PDB:2PS4"
FT BINDING 241
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000269|PubMed:16171386,
FT ECO:0000269|PubMed:17996718, ECO:0007744|PDB:2AEK,
FT ECO:0007744|PDB:2PS4"
FT MUTAGEN 100
FT /note="D->E: Does not significantly perturb the overall
FT structure of trichodiene synthase but leads to an increased
FT KM, a reduction in kcat, as well as to the production of
FT anomalous sesquiterpene products in addition to trichodiene
FT when incubated with farnesyl diphosphate."
FT /evidence="ECO:0000269|PubMed:11827517,
FT ECO:0000269|PubMed:8823172"
FT MUTAGEN 101
FT /note="D->E: Leads to an increased KM for Mg(2+), a
FT reduction in kcat, as well as to the production of
FT anomalous sesquiterpene products in addition to trichodiene
FT when incubated with farnesyl diphosphate."
FT /evidence="ECO:0000269|PubMed:8823172"
FT MUTAGEN 104
FT /note="D->E: Does not significantly affect the KM and kcat
FT for farnesyl diphosphate."
FT /evidence="ECO:0000269|PubMed:8823172"
FT MUTAGEN 146
FT /note="C->F: Leads to the loss of activity."
FT /evidence="ECO:0000269|PubMed:7873527"
FT MUTAGEN 190
FT /note="C->F: Increases the KM for farnesyl diphosphate by
FT about 1.3-fold and reduces the kcat by about 2000-fold."
FT /evidence="ECO:0000269|PubMed:7873527"
FT MUTAGEN 225
FT /note="N->D: Increases the KM for farnesyl diphosphate by
FT about 6-fold and reduces the kcat by about 28-fold. Leads
FT to complete loss of activity; when associated with S-229."
FT /evidence="ECO:0000269|PubMed:17996718"
FT MUTAGEN 229
FT /note="S->T: Increases the KM for farnesyl diphosphate by
FT about 77-fold and reduces the kcat by about 9-fold. Leads
FT to complete loss of activity; when associated with D-225."
FT /evidence="ECO:0000269|PubMed:17996718"
FT MUTAGEN 295
FT /note="Y->F: Does not affect the catalytic activity."
FT /evidence="ECO:0000269|PubMed:17996718"
FT MUTAGEN 304
FT /note="R->K: Does not cause large changes in the overall
FT structure but increases the KM for farnesyl diphosphate by
FT about 25-fold, reduces the kcat by about 200-fold, and
FT leads to conversion of farnesyl diphosphate not only to
FT trichodiene but to at least 2 additional C(15)H(24)
FT hydrocarbons."
FT /evidence="ECO:0000269|PubMed:16171386,
FT ECO:0000269|PubMed:7873527"
FT MUTAGEN 305
FT /note="Y->F: Does not cause large changes in the overall
FT structure but increases the KM for farnesyl diphosphate by
FT about 7-fold."
FT /evidence="ECO:0000269|PubMed:15835903,
FT ECO:0000269|PubMed:7873527"
FT MUTAGEN 305
FT /note="Y->T: Increases the KM for farnesyl diphosphate by
FT about 80-fold, reduces the kcat by about 120-fold, and
FT leads to the conversion of farneyl diphosphate to an
FT approximately equal mixture of trichodiene and an
FT unidentified sesquiterpene hydrocarbon."
FT /evidence="ECO:0000269|PubMed:7873527"
FT HELIX 6..19
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 29..47
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 50..55
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 60..77
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 83..99
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 107..110
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 113..119
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 126..139
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 144..164
FT /evidence="ECO:0007829|PDB:1YYQ"
FT TURN 165..167
FT /evidence="ECO:0007829|PDB:2PS4"
FT HELIX 177..186
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 188..193
FT /evidence="ECO:0007829|PDB:1YYQ"
FT TURN 198..200
FT /evidence="ECO:0007829|PDB:1YYQ"
FT TURN 203..206
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 207..233
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 243..251
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 255..277
FT /evidence="ECO:0007829|PDB:1YYQ"
FT STRAND 278..280
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 282..301
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 303..305
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 308..314
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 320..336
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 340..342
FT /evidence="ECO:0007829|PDB:1YYQ"
FT HELIX 348..353
FT /evidence="ECO:0007829|PDB:1YYQ"
SQ SEQUENCE 374 AA; 44000 MW; 189E8FC1663C3763 CRC64;
MENFPTEYFL NTTVRLLEYI RYRDSNYTRE ERIENLHYAY NKAAHHFAQP RQQQLLKVDP
KRLQASLQTI VGMVVYSWAK VSKECMADLS IHYTYTLVLD DSKDDPYPTM VNYFDDLQAG
REQAHPWWAL VNEHFPNVLR HFGPFCSLNL IRSTLDFFEG CWIEQYNFGG FPGSHDYPQF
LRRMNGLGHC VGASLWPKEQ FNERSLFLEI TSAIAQMENW MVWVNDLMSF YKEFDDERDQ
ISLVKNYVVS DEISLHEALE KLTQDTLHSS KQMVAVFSDK DPQVMDTIEC FMHGYVTWHL
CDRRYRLSEI YEKVKEEKTE DAQKFCKFYE QAANVGAVSP SEWAYPPVAQ LANVRSKDVK
EVQKPFLSSI ELVE
