TPS8_PSEAD
ID TPS8_PSEAD Reviewed; 846 AA.
AC A0A1L6Z3A0;
DT 17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT 15-MAR-2017, sequence version 1.
DT 03-AUG-2022, entry version 21.
DE RecName: Full=Pseudolaratriene synthase, chloroplastic {ECO:0000303|PubMed:28096378};
DE EC=4.2.3.180 {ECO:0000269|PubMed:28096378};
DE AltName: Full=Terpene synthase 8 {ECO:0000303|PubMed:28096378};
DE Short=PxaTPS8 {ECO:0000303|PubMed:28096378};
DE Flags: Precursor;
GN Name=TPS8 {ECO:0000303|PubMed:28096378};
OS Pseudolarix amabilis (Golden larch) (Pseudolarix kaempferi).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Pinopsida; Pinidae; Conifers I; Pinales; Pinaceae;
OC Pseudolarix.
OX NCBI_TaxID=3355;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR, TISSUE
RP SPECIFICITY, AND MUTAGENESIS OF ARG-558; TYR-564; PHE-584; ALA-591;
RP HIS-670; SER-696; GLY-697; ALA-701; GLY-735 AND ALA-743.
RX PubMed=28096378; DOI=10.1073/pnas.1612901114;
RA Mafu S., Karunanithi P.S., Palazzo T.A., Harrod B.L., Rodriguez S.M.,
RA Mollhoff I.N., O'Brien T.E., Tong S., Fiehn O., Tantillo D.J., Bohlmann J.,
RA Zerbe P.;
RT "Biosynthesis of the microtubule-destabilizing diterpene pseudolaric acid B
RT from golden larch involves an unusual diterpene synthase.";
RL Proc. Natl. Acad. Sci. U.S.A. 114:974-979(2017).
CC -!- FUNCTION: Converts geranylgeranyl diphosphate to an new 5,7-fused
CC bicyclic diterpene, named pseudolaratriene (PubMed:28096378). Catalyzes
CC the first committed step in pseudolaric acid B (PAB) biosynthesis
CC (PubMed:28096378). PAB exhibits antiproliferative activity by
CC inhibiting microtubule polymerization, and has demonstrated antitumor
CC properties against several cancer types (Probable).
CC {ECO:0000269|PubMed:28096378, ECO:0000305|PubMed:28096378}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E,10E)-geranylgeranyl diphosphate = diphosphate +
CC pseudolaratriene; Xref=Rhea:RHEA:54116, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:58756, ChEBI:CHEBI:138050; EC=4.2.3.180;
CC Evidence={ECO:0000269|PubMed:28096378};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:54117;
CC Evidence={ECO:0000269|PubMed:28096378};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000305|PubMed:28096378};
CC -!- PATHWAY: Terpene metabolism. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: Plastid, chloroplast {ECO:0000255}.
CC -!- TISSUE SPECIFICITY: Expressed in young and mature roots
CC (PubMed:28096378). Expressed at low levels in barks (PubMed:28096378).
CC {ECO:0000269|PubMed:28096378}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; KU685114; APT40486.1; -; mRNA.
DR AlphaFoldDB; A0A1L6Z3A0; -.
DR SMR; A0A1L6Z3A0; -.
DR KEGG; ag:APT40486; -.
DR BRENDA; 4.2.3.180; 15341.
DR GO; GO:0009507; C:chloroplast; IEA:UniProtKB-SubCell.
DR GO; GO:0000287; F:magnesium ion binding; IEA:InterPro.
DR GO; GO:0010333; F:terpene synthase activity; IEA:InterPro.
DR GO; GO:0016102; P:diterpenoid biosynthetic process; IEA:InterPro.
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; IPR034741; Terpene_cyclase-like_1_C.
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 SFLD; SFLDG01019; Terpene_Cyclase_Like_1_C_Termi; 1.
DR SUPFAM; SSF48239; SSF48239; 2.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW Chloroplast; Lyase; Magnesium; Metal-binding; Plastid; Transit peptide.
FT TRANSIT 1..58
FT /note="Chloroplast"
FT /evidence="ECO:0000255"
FT CHAIN 59..846
FT /note="Pseudolaratriene synthase, chloroplastic"
FT /id="PRO_0000450341"
FT MOTIF 595..599
FT /note="DDXXD motif"
FT /evidence="ECO:0000305"
FT BINDING 595
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:O81086"
FT BINDING 595
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:O81086"
FT BINDING 599
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:O81086"
FT BINDING 599
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:O81086"
FT BINDING 747
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:O81086"
FT MUTAGEN 558
FT /note="R->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 564
FT /note="Y->F: No effect on catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 564
FT /note="Y->H: Slightly reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 564
FT /note="Y->I,T,V: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 564
FT /note="Y->W: Almost abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 584
FT /note="F->Y: Slightly reduces catalytic activity."
FT MUTAGEN 591
FT /note="A->Q: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 670
FT /note="H->A,W,Y: Slightly reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 696
FT /note="S->I,V: Slightly reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 697
FT /note="G->S: Strongly reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 701
FT /note="A->C: Reduces catalytic activity 2-fold."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 701
FT /note="A->L: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 735
FT /note="G->D,N,W: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 735
FT /note="G->S: Slightly reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
FT MUTAGEN 743
FT /note="A->T: Strongly reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:28096378"
SQ SEQUENCE 846 AA; 96588 MW; 3F010A445914E071 CRC64;
MSRFTSATHG LNLSIKMPIS VSQVPSIRSN TSKYELQKLR STGRSVLQTR RQLAIINMTK
RSEADDNDGV ERRKGVFHPN LWDDGFIQSL STVYHEQASY RERAERLIGE VKAVFDSISM
GDGDQFISPS AYDTAWVARV PAIDGSSRPQ FPQAIDWILL NQQQDGSWGS QSHLSLTHRL
TDTLACVIAL ASWKIESVQI DEGLDFITRG VEKLQSESVP AEFEIIFAEL LNQAKSLQLS
LPYEHSCLQS LWRKQEPILA NGLMDSVAKR SLSSLEEMQD HRMNTDSDGT MHVESFLSSP
AVAARVLMRT GNPICLAYLN NVLNKFGDYV PGMYPVDLFQ RLWMVDNVER LGIDRHFKKE
IQVTLDYVYS YWNGKGIGCG RDSLSPDLNS TSLGFRTLRL HGYNVSADVL EHFKDRDGKF
VCSSNPTVGE IRSVLNLYRA SLLAFPGEKV MEEAETFARR YLEEIVQKIP PSKFSREIEY
VLEFGWQSTV PRWEARSYID FHGLDTYSPW TIYEMASEKF LELAKLEFNI FNSLQHTELQ
YLSRWWNDSG MSQMRFTRHR NVEYYTMASC IAMEPSQSAF RIGFTKLCGI ATCIDDIYDT
YGTIDELKLF REAVKRWDPS AIESLPEYMK SVYMVLYELV NEMAQDTERT QGRDTLDYAR
NAWEAIIDAH LVEAEWIASG HIPTFEEYLE NSKVTSGLHI AILPILTLDV PLPDQLPLQE
IDTLSRFHHL ASTIGRLSGD MNAYKIDLAH GEESSCISCY MKDNPGTTEG DAHNYANVTI
SYLMKELNLE LMGQHNRVSF LRTSKKPAFD IYRASNYMYK YRDGYTIADK ETKNLVMRTL
VQAVSL
