COTB2_STRMJ
ID COTB2_STRMJ Reviewed; 307 AA.
AC C9K1X5;
DT 29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT 24-NOV-2009, sequence version 1.
DT 03-AUG-2022, entry version 32.
DE RecName: Full=Cyclooctat-9-en-7-ol synthase {ECO:0000303|PubMed:19635410};
DE EC=4.2.3.146 {ECO:0000269|PubMed:19635410, ECO:0000269|PubMed:24914964};
GN Name=CotB2 {ECO:0000303|PubMed:19635410};
OS Streptomyces melanosporofaciens.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces; Streptomyces violaceusniger group.
OX NCBI_TaxID=67327 {ECO:0000312|EMBL:BAI44338.1};
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], CATALYTIC ACTIVITY, FUNCTION, AND
RP SUBUNIT.
RC STRAIN=MI614-43F2 {ECO:0000312|EMBL:BAI44338.1};
RX PubMed=19635410; DOI=10.1016/j.chembiol.2009.06.007;
RA Kim S.Y., Zhao P., Igarashi M., Sawa R., Tomita T., Nishiyama M.,
RA Kuzuyama T.;
RT "Cloning and heterologous expression of the cyclooctatin biosynthetic gene
RT cluster afford a diterpene cyclase and two p450 hydroxylases.";
RL Chem. Biol. 16:736-743(2009).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.64 ANGSTROMS), CATALYTIC ACTIVITY, FUNCTION,
RP SUBUNIT, AND MUTAGENESIS OF PHE-107; ASP-110; ASP-111; ASP-113; PHE-149 AND
RP TRP-288.
RX PubMed=24914964; DOI=10.1107/s1399004714005513;
RA Janke R., Goerner C., Hirte M., Brueck T., Loll B.;
RT "The first structure of a bacterial diterpene cyclase: CotB2.";
RL Acta Crystallogr. D 70:1528-1537(2014).
CC -!- FUNCTION: Catalyzes the cyclization of the linear isoprenoid
CC intermediate geranylgeranyl diphosphate to tricycclic cyclooctat-9-en-
CC 7-ol in the cyclooctatin biosynthesis pathway. Cyclooctatin is a potent
CC inhibitor of lysophospholipase. {ECO:0000269|PubMed:19635410,
CC ECO:0000269|PubMed:24914964}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=geranylgeranyl diphosphate + H2O = cyclooctat-9-en-7-ol +
CC diphosphate; Xref=Rhea:RHEA:41628, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:57533, ChEBI:CHEBI:78352;
CC EC=4.2.3.146; Evidence={ECO:0000269|PubMed:19635410,
CC ECO:0000269|PubMed:24914964};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:Q40577};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000250|UniProtKB:Q40577};
CC -!- SUBUNIT: Homodimer. {ECO:0000303|PubMed:19635410,
CC ECO:0000303|PubMed:24914964}.
CC -!- DOMAIN: The Asp-Asp-Xaa-Xaa-Asp/Glu(DDXXD/E) motifs is important for
CC the catalytic activity, presumably through binding to Mg(2+).
CC {ECO:0000250}.
CC -!- MISCELLANEOUS: 3,7,18-dolabellatriene produced by the Trp288Gly mutant
CC has potential antibiotic activity against multidrug-resistant S.aureus
CC (MRSA). {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; AB448947; BAI44338.1; -; Genomic_DNA.
DR PDB; 4OMG; X-ray; 1.64 A; A/B=1-307.
DR PDB; 4OMH; X-ray; 1.64 A; A/B=1-307.
DR PDB; 5GUC; X-ray; 1.80 A; A/B=1-307.
DR PDB; 5GUE; X-ray; 1.80 A; A/B/C/D=1-307.
DR PDB; 6GGI; X-ray; 1.80 A; A/B=1-307.
DR PDB; 6GGJ; X-ray; 2.10 A; A/B=1-307.
DR PDB; 6GGK; X-ray; 2.15 A; A/B=1-293.
DR PDB; 6GGL; X-ray; 1.90 A; A/B=1-307.
DR PDB; 7AO0; X-ray; 1.93 A; A/B=1-307.
DR PDB; 7AO1; X-ray; 1.80 A; A/B=1-307.
DR PDB; 7AO2; X-ray; 1.75 A; A/B=1-307.
DR PDB; 7AO3; X-ray; 1.45 A; A/B=1-307.
DR PDB; 7AO4; X-ray; 2.20 A; A/B=1-307.
DR PDB; 7AO5; X-ray; 2.06 A; A/B=1-307.
DR PDBsum; 4OMG; -.
DR PDBsum; 4OMH; -.
DR PDBsum; 5GUC; -.
DR PDBsum; 5GUE; -.
DR PDBsum; 6GGI; -.
DR PDBsum; 6GGJ; -.
DR PDBsum; 6GGK; -.
DR PDBsum; 6GGL; -.
DR PDBsum; 7AO0; -.
DR PDBsum; 7AO1; -.
DR PDBsum; 7AO2; -.
DR PDBsum; 7AO3; -.
DR PDBsum; 7AO4; -.
DR PDBsum; 7AO5; -.
DR AlphaFoldDB; C9K1X5; -.
DR SMR; C9K1X5; -.
DR KEGG; ag:BAI44338; -.
DR BioCyc; MetaCyc:MON-18586; -.
DR BRENDA; 4.2.3.146; 13958.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
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; Isomerase; Lyase; Magnesium; Metal-binding.
FT CHAIN 1..307
FT /note="Cyclooctat-9-en-7-ol synthase"
FT /id="PRO_0000430746"
FT MOTIF 110..113
FT /note="DDXXD motif; degenerate"
FT MOTIF 220..228
FT /note="NSE/DTE motif"
FT BINDING 110
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 110
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 220
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 224
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT BINDING 228
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q40577"
FT MUTAGEN 107
FT /note="F->A,G: Produces R-cembrene-A."
FT /evidence="ECO:0000269|PubMed:24914964"
FT MUTAGEN 110
FT /note="D->E: No change in product (cyclooctat-9-en-7-ol)."
FT /evidence="ECO:0000269|PubMed:24914964"
FT MUTAGEN 111
FT /note="D->E: Abolishes activity, no product."
FT /evidence="ECO:0000269|PubMed:24914964"
FT MUTAGEN 113
FT /note="D->E: No change in product (cyclooctat-9-en-7-ol)."
FT /evidence="ECO:0000269|PubMed:24914964"
FT MUTAGEN 149
FT /note="F->G,H,L,V: Produces cyclooctat-9-en-7-ol."
FT /evidence="ECO:0000303|PubMed:24914964"
FT MUTAGEN 149
FT /note="F->Y: Abolishes activity, no product."
FT /evidence="ECO:0000303|PubMed:24914964"
FT MUTAGEN 288
FT /note="W->G: Produces 3,7,18-dolabellatriene."
FT /evidence="ECO:0000269|PubMed:24914964"
FT HELIX 13..15
FT /evidence="ECO:0007829|PDB:5GUC"
FT HELIX 19..21
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 22..36
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 48..62
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 63..65
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 71..84
FT /evidence="ECO:0007829|PDB:7AO3"
FT TURN 87..89
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 93..111
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 114..116
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 119..133
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 136..155
FT /evidence="ECO:0007829|PDB:7AO3"
FT TURN 156..158
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 160..168
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 170..180
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 183..194
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 197..205
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 207..230
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 237..240
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 246..267
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 272..291
FT /evidence="ECO:0007829|PDB:7AO3"
FT HELIX 293..295
FT /evidence="ECO:0007829|PDB:7AO3"
FT STRAND 297..299
FT /evidence="ECO:0007829|PDB:7AO3"
FT TURN 301..303
FT /evidence="ECO:0007829|PDB:7AO3"
SQ SEQUENCE 307 AA; 35445 MW; B04930110A449560 CRC64;
MTTGLSTAGA QDIGRSSVRP YLEECTRRFQ EMFDRHVVTR PTKVELTDAE LREVIDDCNA
AVAPLGKTVS DERWISYVGV VLWSQSPRHI KDMEAFKAVC VLNCVTFVWD DMDPALHDFG
LFLPQLRKIC EKYYGPEDAE VAYEAARAFV TSDHMFRDSP IKAALCTTSP EQYFRFRVTD
IGVDFWMKMS YPIYRHPEFT EHAKTSLAAR MTTRGLTIVN DFYSYDREVS LGQITNCFRL
CDVSDETAFK EFFQARLDDM IEDIECIKAF DQLTQDVFLD LIYGNFVWTT SNKRYKTAVN
DVNSRIQ
