EASH_CLAP2
ID EASH_CLAP2 Reviewed; 314 AA.
AC G8GV69; M1VVW4;
DT 15-MAR-2017, integrated into UniProtKB/Swiss-Prot.
DT 25-JAN-2012, sequence version 1.
DT 03-AUG-2022, entry version 30.
DE RecName: Full=Dioxygenase easH {ECO:0000303|PubMed:24361048};
DE EC=1.14.11.- {ECO:0000269|PubMed:24361048};
DE AltName: Full=Ergot alkaloid biosynthesis protein H {ECO:0000303|PubMed:24361048};
GN Name=easH {ECO:0000303|PubMed:24361048}; Synonyms=easH1, orfC;
GN ORFNames=CPUR_04075;
OS Claviceps purpurea (strain 20.1) (Ergot fungus) (Sphacelia segetum).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Clavicipitaceae; Claviceps.
OX NCBI_TaxID=1111077;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=20.1;
RA D'Auria J.C., Beuerle T., Pichersky E., Dudareva N.;
RL Submitted (JUN-2011) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=20.1;
RX PubMed=23468653; DOI=10.1371/journal.pgen.1003323;
RA Schardl C.L., Young C.A., Hesse U., Amyotte S.G., Andreeva K., Calie P.J.,
RA Fleetwood D.J., Haws D.C., Moore N., Oeser B., Panaccione D.G.,
RA Schweri K.K., Voisey C.R., Farman M.L., Jaromczyk J.W., Roe B.A.,
RA O'Sullivan D.M., Scott B., Tudzynski P., An Z., Arnaoudova E.G.,
RA Bullock C.T., Charlton N.D., Chen L., Cox M., Dinkins R.D., Florea S.,
RA Glenn A.E., Gordon A., Gueldener U., Harris D.R., Hollin W., Jaromczyk J.,
RA Johnson R.D., Khan A.K., Leistner E., Leuchtmann A., Li C., Liu J., Liu J.,
RA Liu M., Mace W., Machado C., Nagabhyru P., Pan J., Schmid J., Sugawara K.,
RA Steiner U., Takach J.E., Tanaka E., Webb J.S., Wilson E.V., Wiseman J.L.,
RA Yoshida R., Zeng Z.;
RT "Plant-symbiotic fungi as chemical engineers: Multi-genome analysis of the
RT Clavicipitaceae reveals dynamics of alkaloid loci.";
RL PLoS Genet. 9:E1003323-E1003323(2013).
RN [3]
RP BIOTECHNOLOGY.
RC STRAIN=P1 / 1029/N5;
RX PubMed=11778866; DOI=10.1007/s002530100801;
RA Tudzynski P., Correia T., Keller U.;
RT "Biotechnology and genetics of ergot alkaloids.";
RL Appl. Microbiol. Biotechnol. 57:593-605(2001).
RN [4]
RP FUNCTION.
RX PubMed=14700635; DOI=10.1016/j.chembiol.2003.11.013;
RA Correia T., Grammel N., Ortel I., Keller U., Tudzynski P.;
RT "Molecular cloning and analysis of the ergopeptine assembly system in the
RT ergot fungus Claviceps purpurea.";
RL Chem. Biol. 10:1281-1292(2003).
RN [5]
RP FUNCTION.
RC STRAIN=ATCC 20102 / Farmitalia FI 32/17;
RX PubMed=14732265; DOI=10.1016/j.fgb.2003.10.002;
RA Wang J., Machado C., Panaccione D.G., Tsai H.-F., Schardl C.L.;
RT "The determinant step in ergot alkaloid biosynthesis by an endophyte of
RT perennial ryegrass.";
RL Fungal Genet. Biol. 41:189-198(2004).
RN [6]
RP FUNCTION, AND IDENTIFICATION IN THE EAS CLUSTER.
RX PubMed=15904941; DOI=10.1016/j.phytochem.2005.04.011;
RA Haarmann T., Machado C., Lubbe Y., Correia T., Schardl C.L.,
RA Panaccione D.G., Tudzynski P.;
RT "The ergot alkaloid gene cluster in Claviceps purpurea: extension of the
RT cluster sequence and intra species evolution.";
RL Phytochemistry 66:1312-1320(2005).
RN [7]
RP FUNCTION.
RC STRAIN=P1 / 1029/N5;
RX PubMed=16538694; DOI=10.1002/cbic.200500487;
RA Haarmann T., Ortel I., Tudzynski P., Keller U.;
RT "Identification of the cytochrome P450 monooxygenase that bridges the
RT clavine and ergoline alkaloid pathways.";
RL ChemBioChem 7:645-652(2006).
RN [8]
RP FUNCTION.
RX PubMed=17308187; DOI=10.1128/aem.00257-07;
RA Fleetwood D.J., Scott B., Lane G.A., Tanaka A., Johnson R.D.;
RT "A complex ergovaline gene cluster in epichloe endophytes of grasses.";
RL Appl. Environ. Microbiol. 73:2571-2579(2007).
RN [9]
RP FUNCTION.
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 [10]
RP FUNCTION.
RX PubMed=17560817; DOI=10.1016/j.fgb.2007.04.008;
RA Haarmann T., Lorenz N., Tudzynski P.;
RT "Use of a nonhomologous end joining deficient strain (Deltaku70) of the
RT ergot fungus Claviceps purpurea for identification of a nonribosomal
RT peptide synthetase gene involved in ergotamine biosynthesis.";
RL Fungal Genet. Biol. 45:35-44(2008).
RN [11]
RP FUNCTION.
RX PubMed=19139103; DOI=10.1074/jbc.m807168200;
RA Ortel I., Keller U.;
RT "Combinatorial assembly of simple and complex D-lysergic acid alkaloid
RT peptide classes in the ergot fungus Claviceps purpurea.";
RL J. Biol. Chem. 284:6650-6660(2009).
RN [12]
RP FUNCTION.
RX PubMed=20118373; DOI=10.1128/aem.00737-09;
RA Lorenz N., Olsovska J., Sulc M., Tudzynski P.;
RT "Alkaloid cluster gene ccsA of the ergot fungus Claviceps purpurea encodes
RT chanoclavine I synthase, a flavin adenine dinucleotide-containing
RT oxidoreductase mediating the transformation of N-methyl-
RT dimethylallyltryptophan to chanoclavine I.";
RL Appl. Environ. Microbiol. 76:1822-1830(2010).
RN [13]
RP FUNCTION.
RC STRAIN=ATCC 20102 / Farmitalia FI 32/17;
RX PubMed=20735127; DOI=10.1021/ja105785p;
RA Cheng J.Z., Coyle C.M., Panaccione D.G., O'Connor S.E.;
RT "Controlling a structural branch point in ergot alkaloid biosynthesis.";
RL J. Am. Chem. Soc. 132:12835-12837(2010).
RN [14]
RP FUNCTION.
RX PubMed=21409592; DOI=10.1007/s00294-011-0336-4;
RA Goetz K.E., Coyle C.M., Cheng J.Z., O'Connor S.E., Panaccione D.G.;
RT "Ergot cluster-encoded catalase is required for synthesis of chanoclavine-I
RT in Aspergillus fumigatus.";
RL Curr. Genet. 57:201-211(2011).
RN [15]
RP FUNCTION.
RX PubMed=21494745; DOI=10.1039/c0ob01215g;
RA Matuschek M., Wallwey C., Xie X., Li S.M.;
RT "New insights into ergot alkaloid biosynthesis in Claviceps purpurea: an
RT agroclavine synthase EasG catalyses, via a non-enzymatic adduct with
RT reduced glutathione, the conversion of chanoclavine-I aldehyde to
RT agroclavine.";
RL Org. Biomol. Chem. 9:4328-4335(2011).
RN [16] {ECO:0007744|PDB:4NAO}
RP X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) IN COMPLEX WITH IRON, COFACTOR,
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=24361048; DOI=10.1016/j.chembiol.2013.11.008;
RA Havemann J., Vogel D., Loll B., Keller U.;
RT "Cyclolization of D-lysergic acid alkaloid peptides.";
RL Chem. Biol. 21:146-155(2014).
CC -!- FUNCTION: Dioxygenase; part of the gene cluster that mediates the
CC biosynthesis of fungal ergot alkaloid (PubMed:14732265,
CC PubMed:14700635, PubMed:15904941, PubMed:17308187, PubMed:17720822).
CC DmaW catalyzes the first step of ergot alkaloid biosynthesis by
CC condensing dimethylallyl diphosphate (DMAP) and tryptophan to form 4-
CC dimethylallyl-L-tryptophan (PubMed:14732265). The second step is
CC catalyzed by the methyltransferase easF that methylates 4-
CC dimethylallyl-L-tryptophan in the presence of S-adenosyl-L-methionine,
CC resulting in the formation of 4-dimethylallyl-L-abrine (By similarity).
CC The catalase easC and the FAD-dependent oxidoreductase easE then
CC transform 4-dimethylallyl-L-abrine to chanoclavine-I which is further
CC oxidized by easD in the presence of NAD(+), resulting in the formation
CC of chanoclavine-I aldehyde (PubMed:20118373, PubMed:21409592).
CC Agroclavine dehydrogenase easG then mediates the conversion of
CC chanoclavine-I aldehyde to agroclavine via a non-enzymatic adduct
CC reaction: the substrate is an iminium intermediate that is formed
CC spontaneously from chanoclavine-I aldehyde in the presence of
CC glutathione (PubMed:20735127, PubMed:21494745). The presence of easA is
CC not required to complete this reaction (PubMed:21494745). Further
CC conversion of agroclavine to paspalic acid is a two-step process
CC involving oxidation of agroclavine to elymoclavine and of elymoclavine
CC to paspalic acid, the second step being performed by the elymoclavine
CC oxidase cloA (PubMed:16538694, PubMed:17720822). Paspalic acid is then
CC further converted to D-lysergic acid (PubMed:15904941). Ergopeptines
CC are assembled from D-lysergic acid and three different amino acids by
CC the D-lysergyl-peptide-synthetases composed each of a monomudular and a
CC trimodular nonribosomal peptide synthetase subunit (PubMed:14700635,
CC PubMed:15904941). LpsB and lpsC encode the monomodular subunits
CC responsible for D-lysergic acid activation and incorporation into the
CC ergopeptine backbone (PubMed:14700635). LpsA1 and A2 subunits encode
CC the trimodular nonribosomal peptide synthetase assembling the
CC tripeptide portion of ergopeptines (PubMed:14700635). LpsA1 is
CC responsible for formation of the major ergopeptine, ergotamine, and
CC lpsA2 for alpha-ergocryptine, the minor ergopeptine of the total
CC alkaloid mixture elaborated by C.purpurea (PubMed:17560817,
CC PubMed:19139103). D-lysergyl-tripeptides are assembled by the
CC nonribosomal peptide synthetases and released as N-(D-lysergyl-
CC aminoacyl)-lactams (PubMed:24361048). Cyclolization of the D-lysergyl-
CC tripeptides is performed by the Fe(2+)/2-ketoglutarate-dependent
CC dioxygenase easH which introduces a hydroxyl group into N-(D-lysergyl-
CC aminoacyl)-lactam at alpha-C of the aminoacyl residue followed by
CC spontaneous condensation with the terminal lactam carbonyl group
CC (PubMed:24361048). {ECO:0000250|UniProtKB:Q50EL0,
CC ECO:0000269|PubMed:14700635, ECO:0000269|PubMed:14732265,
CC ECO:0000269|PubMed:15904941, ECO:0000269|PubMed:16538694,
CC ECO:0000269|PubMed:17560817, ECO:0000269|PubMed:19139103,
CC ECO:0000269|PubMed:20118373, ECO:0000269|PubMed:20735127,
CC ECO:0000269|PubMed:21409592, ECO:0000269|PubMed:21494745,
CC ECO:0000269|PubMed:24361048, ECO:0000305|PubMed:17308187,
CC ECO:0000305|PubMed:17720822}.
CC -!- COFACTOR:
CC Name=Fe cation; Xref=ChEBI:CHEBI:24875;
CC Evidence={ECO:0000269|PubMed:24361048};
CC -!- PATHWAY: Alkaloid biosynthesis; ergot alkaloid biosynthesis.
CC {ECO:0000269|PubMed:24361048}.
CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:Q4WAW9}.
CC -!- SIMILARITY: Belongs to the PhyH family. {ECO:0000305}.
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DR EMBL; JN186799; AET79182.1; -; Genomic_DNA.
DR EMBL; CAGA01000020; CCE30227.1; -; Genomic_DNA.
DR PDB; 4NAO; X-ray; 1.65 A; A=1-314.
DR PDBsum; 4NAO; -.
DR AlphaFoldDB; G8GV69; -.
DR SMR; G8GV69; -.
DR STRING; 1111077.G8GV69; -.
DR EnsemblFungi; CCE30227; CCE30227; CPUR_04075.
DR VEuPathDB; FungiDB:CPUR_04075; -.
DR eggNOG; ENOG502S7ZW; Eukaryota.
DR OrthoDB; 623398at2759; -.
DR UniPathway; UPA00327; -.
DR Proteomes; UP000016801; Unassembled WGS sequence.
DR GO; GO:0051213; F:dioxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0035835; P:indole alkaloid biosynthetic process; IEA:UniProtKB-UniPathway.
DR InterPro; IPR008775; Phytyl_CoA_dOase.
DR Pfam; PF05721; PhyH; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Dioxygenase; Iron; Metal-binding;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..314
FT /note="Dioxygenase easH"
FT /id="PRO_0000439145"
FT BINDING 141
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:24361048,
FT ECO:0007744|PDB:4NAO"
FT BINDING 143
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:24361048,
FT ECO:0007744|PDB:4NAO"
FT BINDING 217
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:24361048,
FT ECO:0007744|PDB:4NAO"
FT STRAND 13..15
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 20..30
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 31..36
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 41..59
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 76..79
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 84..87
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 90..92
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 95..99
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 101..103
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 105..114
FT /evidence="ECO:0007829|PDB:4NAO"
FT TURN 115..118
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 121..126
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 128..131
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 161..168
FT /evidence="ECO:0007829|PDB:4NAO"
FT TURN 172..174
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 177..180
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 183..185
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 198..201
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 208..212
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 224..226
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 228..237
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 251..255
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 259..264
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 270..272
FT /evidence="ECO:0007829|PDB:4NAO"
FT STRAND 282..287
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 292..295
FT /evidence="ECO:0007829|PDB:4NAO"
FT HELIX 305..307
FT /evidence="ECO:0007829|PDB:4NAO"
SQ SEQUENCE 314 AA; 35431 MW; C76536B286794ADC CRC64;
MTSQHQEHTG TKRFSIQSDP VEIHRAIVED GVAIIEGFLT PEQVQKLNKD VDAPLKADRE
QLKFKADKKD DPHFWLADFI PDHVARVHNL VDFSHCFRHE ILNHELLHKI CRLTFEESGD
YWLGYGAVIE NGPGTTEQKW HRDQPRYPLV KEGPDAPEGM LNFFTALTDF DAETGKTQYI
LGSNKRVELG EPDADHPIEY VGLKPGDTTI VSGKITHRGS DNRSDKMRRA MPIMIIPSIL
TPFDATCHLS RELVETMTPL AQKMICRRSV MIPAPGTVEV KTGIWCVNMR EAGEQIGLKS
NQRAKEDAEA TDAV
