DABA_PSEMU
ID DABA_PSEMU Reviewed; 482 AA.
AC A0A386KZ50;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 05-DEC-2018, sequence version 1.
DT 03-AUG-2022, entry version 10.
DE RecName: Full=Magnesium-dependent glutamate N-prenyltransferase {ECO:0000303|PubMed:30262498};
DE EC=2.5.1.- {ECO:0000269|PubMed:30262498, ECO:0000269|PubMed:32457155};
DE AltName: Full=Domoic acid biosynthesis cluster protein A {ECO:0000303|PubMed:30262498};
DE Short=PmDabA {ECO:0000303|PubMed:30262498};
GN Name=dabA {ECO:0000303|PubMed:30262498};
OS Pseudo-nitzschia multiseries (Marine planktonic diatom) (Nitzschia pungens
OS f. multiseries).
OC Eukaryota; Sar; Stramenopiles; Ochrophyta; Bacillariophyta;
OC Bacillariophyceae; Bacillariophycidae; Bacillariales; Bacillariaceae;
OC Pseudo-nitzschia.
OX NCBI_TaxID=37319;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, PATHWAY,
RP AND INDUCTION BY PHOSPHATE LIMITATION AND CO(2).
RC STRAIN=15091C3;
RX PubMed=30262498; DOI=10.1126/science.aau0382;
RA Brunson J.K., McKinnie S.M.K., Chekan J.R., McCrow J.P., Miles Z.D.,
RA Bertrand E.M., Bielinski V.A., Luhavaya H., Obornik M., Smith G.J.,
RA Hutchins D.A., Allen A.E., Moore B.S.;
RT "Biosynthesis of the neurotoxin domoic acid in a bloom-forming diatom.";
RL Science 361:1356-1358(2018).
RN [2]
RP REVIEW ON NEUROTOXIC ACTIVITY.
RX PubMed=10223631;
RX DOI=10.1002/(sici)1522-7189(199805/08)6:3/4<153::aid-nt16>3.0.co;2-1;
RA Hampson D.R., Manalo J.L.;
RT "The activation of glutamate receptors by kainic acid and domoic acid.";
RL Nat. Toxins 6:153-158(1998).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 46-482 IN COMPLEX WITH MAGNESIUM
RP AND MANGANESE, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP THR-143; TYR-167; ARG-358; GLU-359; HIS-436; GLU-437 AND ARG-447, COFACTOR,
RP AND CATALYTIC ACTIVITY.
RX PubMed=32457155; DOI=10.1073/pnas.2001325117;
RA Chekan J.R., McKinnie S.M.K., Noel J.P., Moore B.S.;
RT "Algal neurotoxin biosynthesis repurposes the terpene cyclase structural
RT fold into an N-prenyltransferase.";
RL Proc. Natl. Acad. Sci. U.S.A. 117:12799-12805(2020).
CC -!- FUNCTION: Magnesium-dependent glutamate N-prenyltransferase: part of
CC the gene cluster that mediates the biosynthesis of domoic acid (DA) and
CC derivatives, natural products with neurochemical activity acting as
CC ionotropic glutamate receptor (iGluR) agonists, thus being neurotoxins
CC causing amnesic shellfish poisoning (ASP) (PubMed:30262498). Catalyzes
CC the conversion of L-glutamic acid (L-Glu) to N-geranyl-L-glutamic acid
CC (NGG) in the presence of geranyl diphosphate (GPP) (PubMed:30262498,
CC PubMed:32457155). Also able to catalyze the formation of farnesyl-L-
CC glutamate from farnesyl diphosphate (FPP) (PubMed:32457155). Cannot use
CC dimethylallyl diphosphate (DMAPP) as substrate (PubMed:32457155).
CC {ECO:0000269|PubMed:30262498, ECO:0000269|PubMed:32457155}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E)-geranyl diphosphate + L-glutamate = diphosphate + N-
CC geranyl-L-glutamate; Xref=Rhea:RHEA:68156, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:58057, ChEBI:CHEBI:172365;
CC Evidence={ECO:0000269|PubMed:30262498, ECO:0000269|PubMed:32457155};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:68157;
CC Evidence={ECO:0000269|PubMed:30262498, ECO:0000269|PubMed:32457155};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:32457155};
CC Note=Can also use Mn(2+) ions as cofactors with a lower efficiency.
CC {ECO:0000269|PubMed:32457155};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.33 uM for geranyl diphosphate {ECO:0000269|PubMed:32457155};
CC KM=21 mM for L-glutamic acid {ECO:0000269|PubMed:32457155};
CC Note=kcat is 4.3 min(-1) with geranyl diphosphate as substrate
CC (PubMed:32457155). kcat is 4.8 min(-1) with L-glutamic acid as
CC substrate (PubMed:32457155). {ECO:0000269|PubMed:32457155};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:30262498}.
CC -!- INDUCTION: Up-regulated under phosphate limitation and by increasing
CC partial pressure of CO(2). {ECO:0000269|PubMed:30262498}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; MH202990; AYD91073.1; -; Genomic_DNA.
DR PDB; 6VKZ; X-ray; 2.10 A; A=46-482.
DR PDB; 6VL0; X-ray; 2.20 A; A=46-482.
DR PDB; 6VL1; X-ray; 2.10 A; A=46-482.
DR PDBsum; 6VKZ; -.
DR PDBsum; 6VL0; -.
DR PDBsum; 6VL1; -.
DR SMR; A0A386KZ50; -.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016740; F:transferase activity; IEA:UniProtKB-KW.
DR GO; GO:0071244; P:cellular response to carbon dioxide; IEP:UniProtKB.
DR GO; GO:0016036; P:cellular response to phosphate starvation; IEP:UniProtKB.
DR Gene3D; 1.10.600.10; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Magnesium; Metal-binding; Transferase.
FT CHAIN 1..482
FT /note="Magnesium-dependent glutamate N-prenyltransferase"
FT /id="PRO_0000454273"
FT BINDING 351
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:32457155,
FT ECO:0007744|PDB:6VKZ, ECO:0007744|PDB:6VL0"
FT BINDING 355
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:32457155,
FT ECO:0007744|PDB:6VKZ, ECO:0007744|PDB:6VL0"
FT BINDING 359
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:32457155,
FT ECO:0007744|PDB:6VKZ, ECO:0007744|PDB:6VL0"
FT BINDING 359
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:32457155,
FT ECO:0007744|PDB:6VKZ, ECO:0007744|PDB:6VL0"
FT BINDING 366
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:32457155,
FT ECO:0007744|PDB:6VKZ, ECO:0007744|PDB:6VL0"
FT MUTAGEN 143
FT /note="T->M: Acquired activity on dimethylallyl diphosphate
FT (DMAPP), but reduced efficiency with geranyl diphosphate
FT (GPP) as substrate."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 167
FT /note="Y->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 167
FT /note="Y->F: Normal activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 167
FT /note="Y->L: Strongly reduced activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 358
FT /note="R->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 359
FT /note="E->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 436
FT /note="H->A: Reduced efficiency with L-glutamic acid (L-
FT Glu) as substrate due to a reduced affinity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 437
FT /note="E->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT MUTAGEN 447
FT /note="R->A: Complete loss of activity."
FT /evidence="ECO:0000269|PubMed:32457155"
FT HELIX 48..58
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 62..64
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 66..78
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 80..82
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 86..93
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 96..103
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 114..119
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 121..125
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 128..130
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 132..138
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 140..147
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 153..173
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 174..177
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 180..182
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 185..204
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 211..220
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 221..227
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 228..231
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 234..237
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 239..241
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 249..262
FT /evidence="ECO:0007829|PDB:6VKZ"
FT TURN 263..266
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 270..291
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 299..309
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 312..320
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 323..325
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 328..332
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 334..354
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 356..362
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 368..375
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 381..408
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 413..438
FT /evidence="ECO:0007829|PDB:6VKZ"
FT STRAND 445..448
FT /evidence="ECO:0007829|PDB:6VKZ"
FT HELIX 451..471
FT /evidence="ECO:0007829|PDB:6VKZ"
SQ SEQUENCE 482 AA; 55647 MW; F95BF7047B27328F CRC64;
MKFATSIVAA IATTGAAFTV IPQKLSHPSQ LNALNTMGSI SSITAESPKE VLSRVQDAGL
TLTNPNDLYW MVDFLKEKYY DNGDYYYPIK TVCDGESIDV KFYCPFEPSL SPHYLELYGS
RDERASIYET TMKKYNRINS EKTSAICTPY SSYGDTQIVA YFYSMMYYIN DQTAHLKLPE
SEIESELIDI LNDDILIYLN EFMSIFEPED AQDLERIWDF LDFYQPYFSK VDGKIVLDEK
YLVRTPSQMP LIKTICEYVS EQFAPSKNIT QVIWEVVRYI KGVKDEIHIR GDKSFTLSLQ
EYDDFRDKVT ASPMAHAVSD LTHERFSYEA YTNPAFMELE NRCSEIITYF NDVCTSDRER
LDEDPFNSVF ILMDLDPSLN FAKSCDVVVE HAYNKMQAFL KLKEEILESA SDEEERLALA
RMIKTREDSL IGYVLHEVCC VEDGYARDHK PLMKAFLEEE ITKSLAEKVK FNPVESESVR
LN
