DMAW_CLAFS
ID DMAW_CLAFS Reviewed; 455 AA.
AC Q12594; A8C7S2;
DT 16-AUG-2004, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 74.
DE RecName: Full=Tryptophan dimethylallyltransferase {ECO:0000303|PubMed:17720822};
DE EC=2.5.1.34 {ECO:0000269|PubMed:1605639, ECO:0000269|PubMed:7488077};
DE AltName: Full=4-dimethylallyltryptophan synthase {ECO:0000303|PubMed:7488077};
DE Short=DMATS {ECO:0000303|PubMed:7488077};
DE AltName: Full=All-trans-hexaprenyl-diphosphate synthase {ECO:0000305};
DE AltName: Full=L-tryptophan dimethylallyl transferase {ECO:0000305};
GN Name=dmaW {ECO:0000303|PubMed:7488077};
OS Claviceps fusiformis (Ergot fungus).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Clavicipitaceae; Claviceps.
OX NCBI_TaxID=40602;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 340-349; 351-360 AND
RP 384-407, CATALYTIC ACTIVITY, AND PATHWAY.
RC STRAIN=ATCC 26245 / DSM 2942 / CBS 164.59;
RX PubMed=7488077; DOI=10.1006/bbrc.1995.2599;
RA Tsai H.-F., Wang H., Gebler J.C., Poulter C.D., Schardl C.L.;
RT "The Claviceps purpurea gene encoding dimethylallyltryptophan synthase, the
RT committed step for ergot alkaloid biosynthesis.";
RL Biochem. Biophys. Res. Commun. 216:119-125(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=ATCC 26245 / DSM 2942 / CBS 164.59;
RX PubMed=17720822; DOI=10.1128/aem.01040-07;
RA Lorenz N., Wilson E.V., Machado C., Schardl C.L., Tudzynski P.;
RT "Comparison of ergot alkaloid biosynthesis gene clusters in Claviceps
RT species indicates loss of late pathway steps in evolution of C.
RT fusiformis.";
RL Appl. Environ. Microbiol. 73:7185-7191(2007).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, SUBUNIT, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=ATCC 26245 / DSM 2942 / CBS 164.59;
RX PubMed=1605639; DOI=10.1016/0003-9861(92)90577-j;
RA Gebler J.C., Poulter C.D.;
RT "Purification and characterization of dimethylallyl tryptophan synthase
RT from Claviceps purpurea.";
RL Arch. Biochem. Biophys. 296:308-313(1992).
CC -!- FUNCTION: Tryptophan dimethylallyltransferase; part of the gene cluster
CC that mediates the biosynthesis of fungal ergot alkaloid
CC (PubMed:7488077, PubMed:17720822, PubMed:1605639). DmaW catalyzes the
CC first step of ergot alkaloid biosynthesis by condensing dimethylallyl
CC diphosphate (DMAP) and tryptophan to form 4-dimethylallyl-L-tryptophan
CC (PubMed:7488077, PubMed:1605639). The second step is catalyzed by the
CC methyltransferase easF that methylates 4-dimethylallyl-L-tryptophan in
CC the presence of S-adenosyl-L-methionine, resulting in the formation of
CC 4-dimethylallyl-L-abrine (By similarity). The catalase easC and the
CC FAD-dependent oxidoreductase easE then transform 4-dimethylallyl-L-
CC abrine to chanoclavine-I which is further oxidized by easD in the
CC presence of NAD(+), resulting in the formation of chanoclavine-I
CC aldehyde (By similarity). Agroclavine dehydrogenase easG then mediates
CC the conversion of chanoclavine-I aldehyde to agroclavine via a non-
CC enzymatic adduct reaction: the substrate is an iminium intermediate
CC that is formed spontaneously from chanoclavine-I aldehyde in the
CC presence of glutathione (By similarity). Further conversion of
CC agroclavine to paspalic acid is a two-step process involving oxidation
CC of agroclavine to elymoclavine and of elymoclavine to paspalic acid,
CC the second step being performed by the elymoclavine oxidase cloA
CC (PubMed:17720822). However, cloA does not encode a functional enzyme
CC indicating that C.fusiformis terminates its ergot alkaloid pathway at
CC elymoclavine (PubMed:17720822). {ECO:0000250|UniProtKB:P0CT20,
CC ECO:0000269|PubMed:1605639, ECO:0000269|PubMed:17720822,
CC ECO:0000269|PubMed:7488077}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=dimethylallyl diphosphate + L-tryptophan = 4-(3-methylbut-2-
CC enyl)-L-tryptophan + diphosphate; Xref=Rhea:RHEA:14173,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57623, ChEBI:CHEBI:57912,
CC ChEBI:CHEBI:58209; EC=2.5.1.34; Evidence={ECO:0000269|PubMed:1605639,
CC ECO:0000269|PubMed:7488077};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=14 uM for dimethylallyl diphosphate (in metal-free EDTA buffer)
CC {ECO:0000269|PubMed:1605639};
CC KM=40 uM for L-tryptophan (in metal-free EDTA buffer)
CC {ECO:0000269|PubMed:1605639};
CC KM=8 uM for dimethylallyl diphosphate (in the presence of 4 mM
CC Ca(2+)) {ECO:0000269|PubMed:1605639};
CC KM=17 uM for L-tryptophan (in the presence of 4 mM Ca(2+))
CC {ECO:0000269|PubMed:1605639};
CC KM=8 uM for dimethylallyl diphosphate (in the presence of 4 mM
CC Mg(2+)) {ECO:0000269|PubMed:1605639};
CC KM=12 uM for L-tryptophan (in the presence of 4 mM Mg(2+))
CC {ECO:0000269|PubMed:1605639};
CC Vmax=215 nmol/min/mg enzyme (in metal-free EDTA buffer)
CC {ECO:0000269|PubMed:1605639};
CC Vmax=504 nmol/min/mg enzyme (in the presence of 4 mM Ca(2+))
CC {ECO:0000269|PubMed:1605639};
CC Vmax=455 nmol/min/mg enzyme (in the presence of 4 mM Mg(2+))
CC {ECO:0000269|PubMed:1605639};
CC -!- PATHWAY: Alkaloid biosynthesis; ergot alkaloid biosynthesis.
CC {ECO:0000269|PubMed:1605639, ECO:0000269|PubMed:7488077}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:1605639}.
CC -!- SIMILARITY: Belongs to the tryptophan dimethylallyltransferase family.
CC {ECO:0000305}.
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DR EMBL; L39640; AAC18893.1; -; Genomic_DNA.
DR EMBL; EU006773; ABV57826.1; -; Genomic_DNA.
DR AlphaFoldDB; Q12594; -.
DR SMR; Q12594; -.
DR SABIO-RK; Q12594; -.
DR UniPathway; UPA00327; -.
DR GO; GO:0004659; F:prenyltransferase activity; IEA:UniProt.
DR GO; GO:0050364; F:tryptophan dimethylallyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0035837; P:ergot alkaloid biosynthetic process; IEA:InterPro.
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 InterPro; IPR017796; Trp_dimethylallylTrfase.
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.
DR TIGRFAMs; TIGR03430; trp_dimet_allyl; 1.
PE 1: Evidence at protein level;
KW Alkaloid metabolism; Direct protein sequencing; Transferase.
FT CHAIN 1..455
FT /note="Tryptophan dimethylallyltransferase"
FT /id="PRO_0000181363"
FT BINDING 79..80
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 88
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 99
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 186
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 188
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 190
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 256
FT /ligand="L-tryptophan"
FT /ligand_id="ChEBI:CHEBI:57912"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 269
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 271
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 273
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 355
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 357
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 421
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
FT BINDING 425
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q50EL0"
SQ SEQUENCE 455 AA; 51858 MW; A913F8E60EBBEB63 CRC64;
MMTKAPATAV YDTLSLLFDF PNQEQRLWWH SIAPMFAAML DTAGHNVHDQ YRHLGIFKKH
IIPFLGVYPA QGKHTWPSVL TRYGIPFELS LNCLDSVVRY TFEPTTEHTG TGDDSYNAFA
ILECIQKLVR IQPGIDMEWF SYFRNELVLN ATESARLGRN DSVNQQPIRT QNKLALDLKG
DRFALKVYLY PHLKSIATGV SSHDLIFNSV RKLSQKHTSI QPSFNVLCDY VASRNDPDSN
AAEAEAGVPA SALRARLLSC DLVDPSKSRI KIYLLEQTVS LTAMEDLWTL GGRRTDSSTL
NGLDMMRELW HLLQIPSGFM KYPESDLKLG EVPDEQLPSM VHYALHPDQP MPEPQVYFTV
FGMSDAGITN ALATFFSRHG WYEMAKKYRV FLEGSFPNHD FESLNYLHTY VSFSYRKNKP
YLSVYLHSFE TGQWPAFSDD PTAFNAFKRC DLSLT
