PYRE3_STRRG
ID PYRE3_STRRG Reviewed; 463 AA.
AC K7QRJ5;
DT 07-OCT-2020, integrated into UniProtKB/Swiss-Prot.
DT 07-OCT-2020, sequence version 2.
DT 03-AUG-2022, entry version 30.
DE RecName: Full=Dialkyldecalin synthase {ECO:0000303|PubMed:25730548};
DE EC=5.5.1.- {ECO:0000269|PubMed:25730548};
DE AltName: Full=FAD-dependent [4+2] cyclase {ECO:0000303|PubMed:29657086};
GN Name=pyrE3 {ECO:0000312|EMBL:AFV71312.1};
OS Streptomyces rugosporus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=295838;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NRRL 21084;
RX PubMed=23062149; DOI=10.1021/ja304829g;
RA Wu Q., Wu Z., Qu X., Liu W.;
RT "Insights into pyrroindomycin biosynthesis reveal a uniform paradigm for
RT tetramate/tetronate formation.";
RL J. Am. Chem. Soc. 134:17342-17345(2012).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP DISRUPTION PHENOTYPE, PATHWAY, REEXAMINATION OF THE N-TERMINAL SEQUENCE,
RP MUTAGENESIS OF GLU-32 AND ASP-275, AND REACTION MECHANISM.
RX PubMed=25730548; DOI=10.1038/nchembio.1769;
RA Tian Z., Sun P., Yan Y., Wu Z., Zheng Q., Zhou S., Zhang H., Yu F., Jia X.,
RA Chen D., Mandi A., Kurtan T., Liu W.;
RT "An enzymatic [4+2] cyclization cascade creates the pentacyclic core of
RT pyrroindomycins.";
RL Nat. Chem. Biol. 11:259-265(2015).
RN [3] {ECO:0007744|PDB:5XGV}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH FAD, FUNCTION,
RP COFACTOR, SUBUNIT, AND MUTAGENESIS OF HIS-74.
RX PubMed=29657086; DOI=10.1016/j.chembiol.2018.03.007;
RA Zheng Q., Gong Y., Guo Y., Zhao Z., Wu Z., Zhou Z., Chen D., Pan L.,
RA Liu W.;
RT "Structural Insights into a Flavin-Dependent [4+2] Cyclase that Catalyzes
RT trans-Decalin Formation in Pyrroindomycin Biosynthesis.";
RL Cell Chem. Biol. 25:718-727.e3(2018).
CC -!- FUNCTION: Involved in the biosynthesis of the spirotetramate
CC antibiotics pyrroindomycins. Catalyzes the intramolecular cyclization
CC forming the dialkyldecalin moiety in pyrroindomycins, via an endo-
CC selective [4+2] cycloaddition reaction. {ECO:0000269|PubMed:25730548,
CC ECO:0000269|PubMed:29657086}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-[(2E,7S,8E,10E,13R,14R,16E,18E)-14-ethyl-7,13-dihydroxy-
CC 2,16,18-trimethylicosa-2,8,10,16,18-pentaenoyl]-2-methylidene-5-oxo-
CC 2,5-dihydro-1H-pyrrol-3-olate = 4-[(1R,2R,4aS,5S,8aR)-2-
CC [(2R,3R,5E,7E)-3-ethyl-2-hydroxy-5,7-dimethylnona-5,7-dien-1-yl]-5-
CC hydroxy-1-methyl-1,2,4a,5,6,7,8,8a-octahydronaphthalene-1-carbonyl]-
CC 2-methylidene-5-oxo-2,5-dihydro-1H-pyrrol-3-olate;
CC Xref=Rhea:RHEA:64476, ChEBI:CHEBI:155854, ChEBI:CHEBI:155855;
CC Evidence={ECO:0000269|PubMed:25730548};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:64477;
CC Evidence={ECO:0000269|PubMed:25730548};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:25730548, ECO:0000269|PubMed:29657086};
CC Note=Binds 1 FAD per subunit. The FAD cofactor does not cycle between
CC oxidized and reduced forms and therefore does not have a typical redox
CC role in the reaction; it plays a structural role essential for PyrE3
CC catalysis, to maintain the requisite geometry of the protein and
CC position the 1,3-diene and dienophile groups through specific
CC interactions with the substrate. {ECO:0000269|PubMed:29657086};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=51.5 uM for 4-[(2E,7S,8E,10E,13R,14R,16E,18E)-14-ethyl-7,13-
CC dihydroxy-2,16,18-trimethylicosa-2,8,10,16,18-pentaenoyl]-2-
CC methylidene-5-oxo-2,5-dihydro-1H-pyrrol-3-olate
CC {ECO:0000269|PubMed:25730548};
CC Note=kcat is 223.2 min(-1). {ECO:0000269|PubMed:25730548};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000269|PubMed:25730548}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:29657086}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene lose the ability to
CC produce pyrroindomycins. {ECO:0000269|PubMed:25730548}.
CC -!- SIMILARITY: Belongs to the PheA/TfdB FAD monooxygenase family.
CC {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AFV71312.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305|PubMed:25730548};
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; JX042309; AFV71312.1; ALT_INIT; Genomic_DNA.
DR PDB; 5XGV; X-ray; 2.10 A; A/B=1-463.
DR PDBsum; 5XGV; -.
DR AlphaFoldDB; K7QRJ5; -.
DR SMR; K7QRJ5; -.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR002938; FAD-bd.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR Pfam; PF01494; FAD_binding_3; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; FAD; Flavoprotein; Isomerase.
FT CHAIN 1..463
FT /note="Dialkyldecalin synthase"
FT /id="PRO_0000450858"
FT BINDING 13
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29657086,
FT ECO:0007744|PDB:5XGV"
FT BINDING 32..33
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29657086,
FT ECO:0007744|PDB:5XGV"
FT BINDING 121
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29657086,
FT ECO:0007744|PDB:5XGV"
FT BINDING 275
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:29657086,
FT ECO:0007744|PDB:5XGV"
FT MUTAGEN 32
FT /note="E->A: Loss of catalytic activity. Loss of FAD
FT binding."
FT /evidence="ECO:0000269|PubMed:25730548"
FT MUTAGEN 74
FT /note="H->A: 81% loss of catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:29657086"
FT MUTAGEN 275
FT /note="D->A: Loss of catalytic activity. Loss of FAD
FT binding."
FT /evidence="ECO:0000269|PubMed:25730548"
FT HELIX 16..23
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 28..31
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 33..36
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 48..55
FT /evidence="ECO:0007829|PDB:5XGV"
FT TURN 56..58
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 59..63
FT /evidence="ECO:0007829|PDB:5XGV"
FT TURN 64..66
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 68..70
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 82..84
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 92..94
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 97..110
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 114..116
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 121..127
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 130..135
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 140..148
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 155..159
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 171..179
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 185..189
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 191..194
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 196..206
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 209..219
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 230..241
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 247..255
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 259..261
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 265..267
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 270..272
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 274..276
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 286..306
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 314..338
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 343..356
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 360..370
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 382..385
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 388..391
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 392..396
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 400..407
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 418..423
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 432..436
FT /evidence="ECO:0007829|PDB:5XGV"
FT STRAND 440..445
FT /evidence="ECO:0007829|PDB:5XGV"
FT HELIX 451..459
FT /evidence="ECO:0007829|PDB:5XGV"
SQ SEQUENCE 463 AA; 49361 MW; 98C84D9B2CC45113 CRC64;
MSDTVIIAGG GPVGLMLACE LGLAGVDTVV LERHDAPREP SRGGAINATV VELFTQRGIM
ESLRDDGFEF RMAHFAHIPL APERVPGDRA FSFAVPHAQV ERRLEERARS LGVRVRRSTE
ITSVRQTPDG VQVTTGDGEV VEGAYLVGCD GSASLVREQA GIPFPGVDPD FHGLWGDIKV
EPGAPVLERI GARQYELGLC MVAPIGPDTV RVITGEFDVP SPPADQEVGF DELRAAVARI
AGVELDGVPG WLSRWTATSR QAERYREGRI LLAGDAAHTL FPLGGQALGT GIEDAVNLGW
KLAATVQGWA PPSLLDSYHE ERHAAGARAC ASTRAQTTIM RSLARVGELR ALLTELAGLE
EVNAYLVRMV GGIDGSRLPD VPLVTAEGET SVYRLLEAGR GVLLDLGAGL PAVRHPQVTY
VRAEPTNRLD ATAVLLRPDG VVAWRAPQDG LEAALETWFG PAA
