SYRB2_PSESY
ID SYRB2_PSESY Reviewed; 310 AA.
AC Q9RBY6;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 61.
DE RecName: Full=L-threonyl-[L-threonyl-carrier protein] 4-chlorinase {ECO:0000305};
DE EC=1.14.20.15 {ECO:0000269|PubMed:16002467, ECO:0000269|PubMed:16528784, ECO:0000269|PubMed:19245217};
DE AltName: Full=Non-haem iron halogenase SyrB2 {ECO:0000303|PubMed:16528784};
DE AltName: Full=Syringomycin biosynthesis enzyme 2 {ECO:0000312|EMBL:AAD50521.1};
DE AltName: Full=Syringomycin synthase SyrB2 {ECO:0000305};
GN Name=syrB2 {ECO:0000303|PubMed:9830033};
OS Pseudomonas syringae pv. syringae.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas; Pseudomonas syringae.
OX NCBI_TaxID=321;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=B301D;
RX PubMed=7608074; DOI=10.1128/jb.177.14.4009-4020.1995;
RA Zhang J.H., Quigley N.B., Gross D.C.;
RT "Analysis of the syrB and syrC genes of Pseudomonas syringae pv. syringae
RT indicates that syringomycin is synthesized by a thiotemplate mechanism.";
RL J. Bacteriol. 177:4009-4020(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], SEQUENCE REVISION, AND PROTEIN SEQUENCE
RP OF 1-7.
RC STRAIN=B301D;
RX PubMed=9830033; DOI=10.1074/jbc.273.49.32857;
RA Guenzi E., Galli G., Grgurina I., Gross D.C., Grandi G.;
RT "Characterization of the syringomycin synthetase gene cluster. A link
RT between prokaryotic and eukaryotic peptide synthetases.";
RL J. Biol. Chem. 273:32857-32863(1998).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND COFACTOR.
RC STRAIN=B301D;
RX PubMed=16002467; DOI=10.1073/pnas.0504412102;
RA Vaillancourt F.H., Yin J., Walsh C.T.;
RT "SyrB2 in syringomycin E biosynthesis is a nonheme FeII alpha-
RT ketoglutarate- and O2-dependent halogenase.";
RL Proc. Natl. Acad. Sci. U.S.A. 102:10111-10116(2005).
RN [4]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=B301D;
RX PubMed=16528784; DOI=10.1002/cbic.200500480;
RA Vaillancourt F.H., Vosburg D.A., Walsh C.T.;
RT "Dichlorination and bromination of a threonyl-S-carrier protein by the non-
RT heme Fe(II) halogenase SyrB2.";
RL ChemBioChem 7:748-752(2006).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, REACTION MECHANISM, AND ACTIVITY REGULATION.
RC STRAIN=B301D;
RX PubMed=19245217; DOI=10.1021/bi900109z;
RA Matthews M.L., Krest C.M., Barr E.W., Vaillancourt F.H., Walsh C.T.,
RA Green M.T., Krebs C., Bollinger J.M.;
RT "Substrate-triggered formation and remarkable stability of the C-H bond-
RT cleaving chloroferryl intermediate in the aliphatic halogenase, SyrB2.";
RL Biochemistry 48:4331-4343(2009).
RN [6]
RP FUNCTION IN NITRATION AND AZIDATION, BIOTECHNOLOGY, AND MUTAGENESIS OF
RP ALA-118.
RX PubMed=24463698; DOI=10.1038/nchembio.1438;
RA Matthews M.L., Chang W.C., Layne A.P., Miles L.A., Krebs C.,
RA Bollinger J.M. Jr.;
RT "Direct nitration and azidation of aliphatic carbons by an iron-dependent
RT halogenase.";
RL Nat. Chem. Biol. 10:209-215(2014).
RN [7] {ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCU, ECO:0007744|PDB:2FCV}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) IN COMPLEXES WITH 2-OXOGLUTARIC
RP ACID; IRON; CHLORIDE ION AND BROMIDE ION, COFACTOR, REACTION MECHANISM,
RP DOMAIN, AND MUTAGENESIS OF ALA-118.
RX PubMed=16541079; DOI=10.1038/nature04544;
RA Blasiak L.C., Vaillancourt F.H., Walsh C.T., Drennan C.L.;
RT "Crystal structure of the non-haem iron halogenase SyrB2 in syringomycin
RT biosynthesis.";
RL Nature 440:368-371(2006).
CC -!- FUNCTION: Involved in the biosynthesis of syringomycin E, a cyclic
CC lipodepsinonapeptide toxin with phytotoxic activity (PubMed:16002467,
CC PubMed:16528784, PubMed:19245217). Catalyzes the chlorination of L-Thr
CC to 4-Cl-L-Thr when the amino acid is tethered via a thioester linkage
CC to a covalently bound phosphopantetheine cofactor of the carrier
CC protein SyrB1 (L-Thr-S-SyrB1), in a reaction that requires oxygen,
CC chloride ions, ferrous iron and 2-oxoglutarate (PubMed:16002467,
CC PubMed:16528784, PubMed:19245217). In vitro, can catalyze tandem
CC chlorinations at the gamma-position of the threonyl-S-protein substrate
CC (PubMed:16528784). Can also brominate L-Thr-S-SyrB1 in reactions
CC performed with excess sodium bromide, but a preference for chloride
CC versus bromide by a factor of 180 can be estimated by comparing the
CC peak intensities (PubMed:16528784). In addition, in the presence of the
CC nitrogenous anions NO2(-) or N3(-), SyrB2 can direct aliphatic
CC nitration and azidation reactions by the same mechanism as the native
CC halogenation reaction (PubMed:24463698). {ECO:0000269|PubMed:16002467,
CC ECO:0000269|PubMed:16528784, ECO:0000269|PubMed:19245217,
CC ECO:0000269|PubMed:24463698}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + chloride + H(+) + L-threonyl-[peptidyl-
CC carrier protein] + O2 = 4-chloro-L-threonyl-[peptidyl-carrier
CC protein] + CO2 + H2O + succinate; Xref=Rhea:RHEA:56812, Rhea:RHEA-
CC COMP:15908, Rhea:RHEA-COMP:15910, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16526,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:17996, ChEBI:CHEBI:30031,
CC ChEBI:CHEBI:144927, ChEBI:CHEBI:144979; EC=1.14.20.15;
CC Evidence={ECO:0000269|PubMed:16002467, ECO:0000269|PubMed:16528784,
CC ECO:0000269|PubMed:19245217};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:56813;
CC Evidence={ECO:0000269|PubMed:16002467, ECO:0000269|PubMed:16528784,
CC ECO:0000269|PubMed:19245217};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:16002467, ECO:0000269|PubMed:16541079};
CC Note=Binds 1 Fe(2+) ion. {ECO:0000269|PubMed:16541079};
CC -!- ACTIVITY REGULATION: The complete trimodular substrate, with carrier
CC protein (SyrB1), covalently attached PPant cofactor, and thioester-
CC appended amino acid, is required for substrate triggering of rapid
CC chloroferryl intermediate formation. {ECO:0000269|PubMed:19245217}.
CC -!- DOMAIN: Iron is coordinated by two histidines and one chloride ligand,
CC in contrast to the two-histidine/one carboxylate facial triad of other
CC mononuclear non-haem iron enzymes. The remaining iron coordination
CC sites are occupied by the 2-oxoglutarate and a water molecule.
CC {ECO:0000269|PubMed:16541079}.
CC -!- BIOTECHNOLOGY: The ability to install nitrogenous functional groups, in
CC particular the azido group, onto aliphatic carbon centers could enable
CC new methods in the field of chemical biology.
CC {ECO:0000269|PubMed:24463698}.
CC -!- SIMILARITY: Belongs to the PhyH family. {ECO:0000305}.
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DR EMBL; U25130; AAD50521.1; -; Genomic_DNA.
DR RefSeq; WP_032655799.1; NZ_QPBU01000111.1.
DR PDB; 2FCT; X-ray; 1.60 A; A/B=1-310.
DR PDB; 2FCU; X-ray; 1.60 A; A/B=1-310.
DR PDB; 2FCV; X-ray; 1.80 A; A/B=1-310.
DR PDBsum; 2FCT; -.
DR PDBsum; 2FCU; -.
DR PDBsum; 2FCV; -.
DR SMR; Q9RBY6; -.
DR BioCyc; MetaCyc:MON-20500; -.
DR BRENDA; 1.14.20.15; 12469.
DR EvolutionaryTrace; Q9RBY6; -.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR InterPro; IPR010092; Chlorin_enz.
DR InterPro; IPR008775; Phytyl_CoA_dOase.
DR Pfam; PF05721; PhyH; 1.
DR TIGRFAMs; TIGR01762; chlorin-enz; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Iron; Metal-binding;
KW Oxidoreductase.
FT CHAIN 1..310
FT /note="L-threonyl-[L-threonyl-carrier protein] 4-
FT chlorinase"
FT /id="PRO_0000454787"
FT BINDING 113..116
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCU,
FT ECO:0007744|PDB:2FCV"
FT BINDING 116
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCV"
FT BINDING 145
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCU,
FT ECO:0007744|PDB:2FCV"
FT BINDING 235
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCU,
FT ECO:0007744|PDB:2FCV"
FT BINDING 235
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCV"
FT BINDING 248
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCU,
FT ECO:0007744|PDB:2FCV"
FT BINDING 254
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:16541079,
FT ECO:0007744|PDB:2FCT, ECO:0007744|PDB:2FCU,
FT ECO:0007744|PDB:2FCV"
FT MUTAGEN 118
FT /note="A->D,E: Abrogates chlorination activity. Can still
FT bind iron."
FT /evidence="ECO:0000269|PubMed:16541079"
FT MUTAGEN 118
FT /note="A->G: Enhances C-N coupling reactions by diminishing
FT affinity for the native Cl(-). Enhance the yield of
FT nitration by 2.5-fold and the yield of azidation by 13-fold
FT in the absence of added Cl(-)."
FT /evidence="ECO:0000269|PubMed:24463698"
FT HELIX 9..18
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 19..27
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 29..43
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 54..56
FT /evidence="ECO:0007829|PDB:2FCU"
FT HELIX 62..65
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 68..70
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 73..78
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 82..92
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 96..106
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 119..121
FT /evidence="ECO:0007829|PDB:2FCT"
FT TURN 123..125
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 129..131
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 142..149
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 158..161
FT /evidence="ECO:0007829|PDB:2FCT"
FT TURN 180..182
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 191..195
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 200..203
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 204..206
FT /evidence="ECO:0007829|PDB:2FCT"
FT HELIX 213..215
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 217..219
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 225..230
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 233..237
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 242..246
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 248..257
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 260..263
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 267..271
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 274..277
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 283..287
FT /evidence="ECO:0007829|PDB:2FCT"
FT STRAND 306..308
FT /evidence="ECO:0007829|PDB:2FCT"
SQ SEQUENCE 310 AA; 35360 MW; 8B07D49BB68DCE54 CRC64;
MSKKFALTAE QRASFEKNGF IGPFDAYSPE EMKETWKRTR LRLLDRSAAA YQDLDAISGG
TNIANYDRHL DDDFLASHIC RPEICDRVES ILGPNVLCWR TEFFPKYPGD EGTDWHQADT
FANASGKPQI IWPENEEFGG TITVWTAFTD ANIANGCLQF IPGTQNSMNY DETKRMTYEP
DANNSVVKDG VRRGFFGYDY RQLQIDENWK PDEASAVPMQ MKAGQFIIFW STLMHASYPH
SGESQEMRMG FASRYVPSFV HVYPDSDHIE EYGGRISLEK YGAVQVIGDE TPEYNRLVTH
TTRGKKFEAV
