PYRI4_STRRG
ID PYRI4_STRRG Reviewed; 184 AA.
AC K7QVW7;
DT 07-OCT-2020, integrated into UniProtKB/Swiss-Prot.
DT 07-OCT-2020, sequence version 2.
DT 03-AUG-2022, entry version 25.
DE RecName: Full=Spiro-conjugate synthase {ECO:0000303|PubMed:25730548, ECO:0000303|PubMed:26877021};
DE EC=5.5.1.- {ECO:0000269|PubMed:25730548};
DE AltName: Full=[4+2] cyclase PyrI4 {ECO:0000303|PubMed:25730548};
GN Name=pyrI4 {ECO:0000312|EMBL:AFV71338.1};
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, BIOPHYSICOCHEMICAL PROPERTIES, DISRUPTION
RP PHENOTYPE, PATHWAY, AND REEXAMINATION OF THE N-TERMINAL SEQUENCE.
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:5BTU, ECO:0007744|PDB:5BU3}
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH ITS
RP PRODUCT, DISULFIDE BOND, FUNCTION, REACTION MECHANISM, SUBUNIT, DOMAIN, AND
RP MUTAGENESIS OF ARG-9; GLU-20; ASP-74 AND GLN-115.
RX PubMed=26877021; DOI=10.1016/j.chembiol.2016.01.005;
RA Zheng Q., Guo Y., Yang L., Zhao Z., Wu Z., Zhang H., Liu J., Cheng X.,
RA Wu J., Yang H., Jiang H., Pan L., Liu W.;
RT "Enzyme-Dependent [4+2] Cycloaddition Depends on Lid-like Interaction of
RT the N-Terminal Sequence with the Catalytic Core in PyrI4.";
RL Cell Chem. Biol. 23:352-360(2016).
CC -!- FUNCTION: Involved in the biosynthesis of the spirotetramate
CC antibiotics pyrroindomycins. Catalyzes the intramolecular cyclization
CC forming the spiro-conjugate moiety in pyrroindomycins, via an exo-
CC selective [4+2] cycloaddition reaction. {ECO:0000269|PubMed:25730548,
CC ECO:0000269|PubMed:26877021}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-[(1R,2R,4aS,5S,8aR)-2-[(2R,3R,5E,7E)-3-ethyl-2-hydroxy-5,7-
CC dimethylnona-5,7-dien-1-yl]-5-hydroxy-1-methyl-1,2,4a,5,6,7,8,8a-
CC octahydronaphthalene-1-carbonyl]-2-methylidene-5-oxo-2,5-dihydro-1H-
CC pyrrol-3-olate = (1S,3R,6R,8R,9R,11R,14S,15S,19R,20R)-8-ethyl-9,15-
CC dihydroxy-3,4,6,20-tetramethyl-21,23-dioxo-24-
CC azapentacyclo[20.2.1.0(1,6).0(11,20).0(14,19)]pentacosa-4,12,22(25)-
CC trien-25-olate; Xref=Rhea:RHEA:64480, ChEBI:CHEBI:155855,
CC ChEBI:CHEBI:155856; Evidence={ECO:0000269|PubMed:25730548};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:64481;
CC Evidence={ECO:0000269|PubMed:25730548};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=224 uM for 4-[(1R,2R,4aS,5S,8aR)-2-[(2R,3R,5E,7E)-3-ethyl-2-
CC hydroxy-5,7-dimethylnona-5,7-dien-1-yl]-5-hydroxy-1-methyl-
CC 1,2,4a,5,6,7,8,8a-octahydronaphthalene-1-carbonyl]-2-methylidene-5-
CC oxo-2,5-dihydro-1H-pyrrol-3-olate {ECO:0000269|PubMed:25730548};
CC Note=kcat is 342.6 min(-1). {ECO:0000269|PubMed:25730548};
CC -!- PATHWAY: Antibiotic biosynthesis. {ECO:0000269|PubMed:25730548}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:26877021}.
CC -!- DOMAIN: The N-terminal 10-AA residues are absolutely required for
CC enzymatic activity. {ECO:0000269|PubMed:26877021}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene lose the ability to
CC produce pyrroindomycins. {ECO:0000269|PubMed:25730548}.
CC -!- MISCELLANEOUS: Reaction occurs via a unique trapping mechanism whereby
CC the lid-like action of the N-terminal tail imposes conformational
CC constraints on the beta-barrel catalytic core, which enhances the
CC proximity and polarization effects of reactive groups (1,3-diene and
CC alkene) to drive cyclization in a regio- and stereo-specific manner.
CC {ECO:0000269|PubMed:26877021}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AFV71338.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305|PubMed:25730548};
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DR EMBL; JX042309; AFV71338.1; ALT_INIT; Genomic_DNA.
DR PDB; 5BTU; X-ray; 2.50 A; A/B=1-184.
DR PDB; 5BU3; X-ray; 1.90 A; A/B/C/D=1-184.
DR PDB; 7DVK; X-ray; 2.60 A; A/B/C/D=11-184.
DR PDBsum; 5BTU; -.
DR PDBsum; 5BU3; -.
DR PDBsum; 7DVK; -.
DR AlphaFoldDB; K7QVW7; -.
DR SMR; K7QVW7; -.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR InterPro; IPR041013; AOC-like.
DR Pfam; PF18678; AOC_like; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Disulfide bond; Isomerase.
FT CHAIN 1..184
FT /note="Spiro-conjugate synthase"
FT /id="PRO_0000450857"
FT BINDING 115
FT /ligand="(1S,3R,6R,8R,9R,11R,14S,15S,19R,20R)-8-ethyl-9,15-
FT dihydroxy-3,4,6,20-tetramethyl-21,23-dioxo-24-
FT azapentacyclo[20.2.1.0(1,6).0(11,20).0(14,19)]pentacosa-
FT 4,12,22(25)-trien-25-olate"
FT /ligand_id="ChEBI:CHEBI:155856"
FT /evidence="ECO:0000269|PubMed:26877021"
FT DISULFID 57..184
FT /evidence="ECO:0007744|PDB:5BTU"
FT MUTAGEN 9
FT /note="R->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:26877021"
FT MUTAGEN 20
FT /note="E->R: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:26877021"
FT MUTAGEN 74
FT /note="D->R: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:26877021"
FT MUTAGEN 115
FT /note="Q->A: Leads to a 60% loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:26877021"
FT HELIX 13..18
FT /evidence="ECO:0007829|PDB:5BU3"
FT HELIX 30..40
FT /evidence="ECO:0007829|PDB:5BU3"
FT HELIX 45..48
FT /evidence="ECO:0007829|PDB:5BTU"
FT HELIX 50..56
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 57..73
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 82..90
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 96..108
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 110..112
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 114..123
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 126..135
FT /evidence="ECO:0007829|PDB:5BU3"
FT HELIX 136..140
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 145..154
FT /evidence="ECO:0007829|PDB:5BU3"
FT TURN 155..158
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 160..169
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 172..174
FT /evidence="ECO:0007829|PDB:5BU3"
FT STRAND 176..184
FT /evidence="ECO:0007829|PDB:5BU3"
SQ SEQUENCE 184 AA; 19481 MW; AA8DC773B090D51D CRC64;
MTTPQIDERA MEAGAAALQE TIVDPGPLDV TALAVAAALA AGLHSAADDP AAALDKCIVL
DELTEFAEKL VVHDRPGGIG TTVEYVEVYE DASGVRLGTA TGNAVVLKME PHMWQFHQSV
SELADGSFEA VGVIDCTAML RRMTQVLRVT GRSGRYAGKS GFMTLAISDP NQRPPHYSVQ
VVLC
