IRIS_CATRO
ID IRIS_CATRO Reviewed; 388 AA.
AC K7WDL7;
DT 29-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 06-FEB-2013, sequence version 1.
DT 03-AUG-2022, entry version 32.
DE RecName: Full=(S)-8-oxocitronellyl enol synthase {ECO:0000305};
DE EC=1.3.1.122 {ECO:0000269|PubMed:23172143, ECO:0000269|PubMed:26551396, ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105};
DE AltName: Full=Iridoid synthase {ECO:0000303|PubMed:30531909};
DE Short=ISY {ECO:0000303|PubMed:30531909};
OS Catharanthus roseus (Madagascar periwinkle) (Vinca rosea).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC asterids; lamiids; Gentianales; Apocynaceae; Rauvolfioideae; Vinceae;
OC Catharanthinae; Catharanthus.
OX NCBI_TaxID=4058;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBCELLULAR LOCATION, TISSUE SPECIFICITY,
RP AND SUBUNIT.
RX PubMed=23172143; DOI=10.1038/nature11692;
RA Geu-Flores F., Sherden N.H., Courdavault V., Burlat V., Glenn W.S., Wu C.,
RA Nims E., Cui Y., O'Connor S.E.;
RT "An alternative route to cyclic terpenes by reductive cyclization in
RT iridoid biosynthesis.";
RL Nature 492:138-142(2012).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=30531909; DOI=10.1038/s41589-018-0185-2;
RA Lichman B.R., Kamileen M.O., Titchiner G.R., Saalbach G., Stevenson C.E.M.,
RA Lawson D.M., O'Connor S.E.;
RT "Uncoupled activation and cyclization in catmint reductive terpenoid
RT biosynthesis.";
RL Nat. Chem. Biol. 15:71-79(2019).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF 26-388 IN COMPLEX WITH NADP AND
RP SUBSTRATE, CATALYTIC ACTIVITY, ACTIVE SITES, AND MUTAGENESIS OF LYS-146;
RP PHE-149; TYR-178; PHE-342; ALA-346 AND SER-349.
RX PubMed=26768532; DOI=10.1002/anie.201508310;
RA Hu Y., Liu W., Malwal S.R., Zheng Y., Feng X., Ko T.P., Chen C.C., Xu Z.,
RA Liu M., Han X., Gao J., Oldfield E., Guo R.T.;
RT "Structures of iridoid synthase from Cantharanthus roseus with bound
RT NAD(+), NADPH, or NAD(+)/10-oxogeranial: reaction mechanisms.";
RL Angew. Chem. Int. Ed. Engl. 54:15478-15482(2015).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) OF 26-388 IN COMPLEX WITH NADP AND
RP SUBSTRATE, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=26868105; DOI=10.1016/j.jsb.2016.02.010;
RA Qin L., Zhu Y., Ding Z., Zhang X., Ye S., Zhang R.;
RT "Structure of iridoid synthase in complex with NADP(+)/8-oxogeranial
RT reveals the structural basis of its substrate specificity.";
RL J. Struct. Biol. 194:224-230(2016).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) OF 23-388 IN COMPLEX WITH NADP,
RP CATALYTIC ACTIVITY, AND SUBUNIT.
RX PubMed=26551396; DOI=10.1038/nchembio.1955;
RA Kries H., Caputi L., Stevenson C.E.M., Kamileen M.O., Sherden N.H.,
RA Geu-Flores F., Lawson D.M., O'Connor S.E.;
RT "Structural determinants of reductive terpene cyclization in iridoid
RT biosynthesis.";
RL Nat. Chem. Biol. 12:6-8(2016).
CC -!- FUNCTION: Iridoid synthase that catalyzes the first step in generation
CC of the iridoid ring scaffold using the linear monoterpene (6E)-8-
CC oxogeranial as substrate. Iridoids comprise a large family of
CC distinctive bicyclic monoterpenes that possess a wide range of
CC pharmacological activities, including anticancer, anti-inflammatory,
CC antifungal and antibacterial activities. {ECO:0000269|PubMed:23172143,
CC ECO:0000269|PubMed:30531909}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-8-oxocitronellyl enol + NADP(+) = (6E)-8-oxogeranial +
CC H(+) + NADPH; Xref=Rhea:RHEA:62592, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:64239,
CC ChEBI:CHEBI:144481; EC=1.3.1.122;
CC Evidence={ECO:0000269|PubMed:23172143, ECO:0000269|PubMed:26768532,
CC ECO:0000269|PubMed:26868105, ECO:0000269|PubMed:30531909};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:62594;
CC Evidence={ECO:0000269|PubMed:23172143, ECO:0000269|PubMed:26768532,
CC ECO:0000269|PubMed:26868105, ECO:0000269|PubMed:30531909};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-8-oxocitronellyl enol + NAD(+) = (6E)-8-oxogeranial + H(+)
CC + NADH; Xref=Rhea:RHEA:62596, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945, ChEBI:CHEBI:64239, ChEBI:CHEBI:144481;
CC EC=1.3.1.122; Evidence={ECO:0000269|PubMed:23172143,
CC ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
CC ECO:0000269|PubMed:30531909};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:62598;
CC Evidence={ECO:0000269|PubMed:23172143, ECO:0000269|PubMed:26768532,
CC ECO:0000269|PubMed:26868105, ECO:0000269|PubMed:30531909};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=4.5 uM for (6E)-8-oxogeranial {ECO:0000269|PubMed:23172143};
CC KM=10.6 uM for (6E)-8-oxogeranial {ECO:0000269|PubMed:26868105};
CC KM=4.7 uM for NADPH {ECO:0000269|PubMed:23172143};
CC Note=kcat is 1.6 sec(-1) with (6E)-8-oxogeranial. kcat is 2.2 sec(-1)
CC with NADPH.;
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:23172143,
CC ECO:0000269|PubMed:26551396, ECO:0000269|PubMed:26768532,
CC ECO:0000269|PubMed:26868105}.
CC -!- INTERACTION:
CC K7WDL7; K7WDL7: -; NbExp=2; IntAct=EBI-16184393, EBI-16184393;
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol {ECO:0000269|PubMed:23172143}.
CC -!- TISSUE SPECIFICITY: Expressed in internal phloem-associated parenchyma
CC (IPAP) cells. {ECO:0000269|PubMed:23172143}.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. Highly divergent. {ECO:0000305}.
CC -!- CAUTION: Was originally thought to catalyze the entire reaction from
CC (6E)-8-oxogeranial to nepetalactol including a cyclase step
CC (PubMed:23172143). New results have shown that the cyclase is a
CC different enzyme and this enzyme exclusively catalyzes a reduction step
CC (PubMed:30531909). {ECO:0000269|PubMed:30531909,
CC ECO:0000305|PubMed:23172143}.
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DR EMBL; JX974564; AFW98981.1; -; mRNA.
DR PDB; 5COA; X-ray; 2.20 A; A/B=26-388.
DR PDB; 5COB; X-ray; 2.65 A; A/B/C/D=26-388.
DR PDB; 5DBF; X-ray; 2.00 A; A/B=26-388.
DR PDB; 5DBG; X-ray; 1.95 A; A/B=26-388.
DR PDB; 5DBI; X-ray; 2.20 A; A/B=26-388.
DR PDB; 5DCU; X-ray; 1.40 A; A/B=23-388.
DR PDB; 5DCW; X-ray; 1.90 A; A=23-388.
DR PDB; 5DCY; X-ray; 1.45 A; A/B=23-388.
DR PDB; 5DF1; X-ray; 1.75 A; A/B=23-388.
DR PDB; 5EMH; X-ray; 2.10 A; A=21-388.
DR PDBsum; 5COA; -.
DR PDBsum; 5COB; -.
DR PDBsum; 5DBF; -.
DR PDBsum; 5DBG; -.
DR PDBsum; 5DBI; -.
DR PDBsum; 5DCU; -.
DR PDBsum; 5DCW; -.
DR PDBsum; 5DCY; -.
DR PDBsum; 5DF1; -.
DR PDBsum; 5EMH; -.
DR AlphaFoldDB; K7WDL7; -.
DR SMR; K7WDL7; -.
DR DIP; DIP-62000N; -.
DR PRIDE; K7WDL7; -.
DR KEGG; ag:AFW98981; -.
DR BioCyc; MetaCyc:MON-20521; -.
DR GO; GO:0005829; C:cytosol; IDA:UniProtKB.
DR GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR GO; GO:0016628; F:oxidoreductase activity, acting on the CH-CH group of donors, NAD or NADP as acceptor; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:0016099; P:monoterpenoid biosynthetic process; IDA:UniProtKB.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; NAD; NADP; Oxidoreductase.
FT CHAIN 1..388
FT /note="(S)-8-oxocitronellyl enol synthase"
FT /id="PRO_0000422594"
FT ACT_SITE 146
FT /evidence="ECO:0000269|PubMed:26768532"
FT ACT_SITE 178
FT /evidence="ECO:0000269|PubMed:26768532"
FT BINDING 38..40
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 66..67
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 84..85
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 108..109
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 142
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 146
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:26768532,
FT ECO:0007744|PDB:5DBI"
FT BINDING 178
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 178
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:26768532,
FT ECO:0000269|PubMed:26868105, ECO:0007744|PDB:5COB,
FT ECO:0007744|PDB:5DBI"
FT BINDING 204
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 211..213
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26551396,
FT ECO:0000269|PubMed:26768532, ECO:0000269|PubMed:26868105,
FT ECO:0007744|PDB:5COB, ECO:0007744|PDB:5DBF,
FT ECO:0007744|PDB:5DF1"
FT BINDING 349
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:26768532,
FT ECO:0000269|PubMed:26868105, ECO:0007744|PDB:5COB"
FT MUTAGEN 146
FT /note="K->A: Reduces enzymatic activity 6-fold."
FT /evidence="ECO:0000269|PubMed:26768532"
FT MUTAGEN 149
FT /note="F->M: Slightly reduces enzymatic activity."
FT /evidence="ECO:0000269|PubMed:26768532"
FT MUTAGEN 178
FT /note="Y->A,F: Abolishes enzymatic activity."
FT /evidence="ECO:0000269|PubMed:26768532"
FT MUTAGEN 342
FT /note="F->A: Abolishes enzymatic activity."
FT /evidence="ECO:0000269|PubMed:26768532"
FT MUTAGEN 346
FT /note="A->I: No effect on enzymatic activity."
FT /evidence="ECO:0000269|PubMed:26768532"
FT MUTAGEN 349
FT /note="S->F: No effect on enzymatic activity."
FT /evidence="ECO:0000269|PubMed:26768532"
FT STRAND 29..36
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 42..48
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 51..54
FT /evidence="ECO:0007829|PDB:5COB"
FT STRAND 58..67
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 71..74
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 79..82
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 88..95
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 103..106
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 111..113
FT /evidence="ECO:0007829|PDB:5COA"
FT HELIX 115..129
FT /evidence="ECO:0007829|PDB:5DCU"
FT TURN 130..132
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 138..142
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 146..149
FT /evidence="ECO:0007829|PDB:5DCU"
FT TURN 153..156
FT /evidence="ECO:0007829|PDB:5DCY"
FT STRAND 163..165
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 177..191
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 195..201
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 203..205
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 215..228
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 239..243
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 248..250
FT /evidence="ECO:0007829|PDB:5DF1"
FT HELIX 251..263
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 265..267
FT /evidence="ECO:0007829|PDB:5DCU"
FT STRAND 270..274
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 282..293
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 309..313
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 317..327
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 334..337
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 340..346
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 357..361
FT /evidence="ECO:0007829|PDB:5DCU"
FT HELIX 370..383
FT /evidence="ECO:0007829|PDB:5DCU"
SQ SEQUENCE 388 AA; 43850 MW; 906C11AD93A04E76 CRC64;
MSWWWKRSIG AGKNLPNQNK ENGVCKSYKS VALVVGVTGI VGSSLAEVLK LPDTPGGPWK
VYGVARRPCP VWLAKKPVEY IQCDVSDNQE TISKLSPLKD ITHIFYVSWI GSEDCQTNAT
MFKNILNSVI PNASNLQHVC LQTGIKHYFG IFEEGSKVVP HDSPFTEDLP RLNVPNFYHD
LEDILYEETG KNNLTWSVHR PALVFGFSPC SMMNIVSTLC VYATICKHEN KALVYPGSKN
SWNCYADAVD ADLVAEHEIW AAVDPKAKNQ VLNCNNGDVF KWKHIWKKLA EEFGIEMVGY
VEGKEQVSLA ELMKDKDQVW DEIVKKNNLV PTKLKEIAAF WFADIAFCSE NLISSMNKSK
ELGFLGFRNS MKSFVSCIDK MRDYRFIP
