RISB2_BRUA2
ID RISB2_BRUA2 Reviewed; 158 AA.
AC Q2YKV1;
DT 19-MAR-2014, integrated into UniProtKB/Swiss-Prot.
DT 20-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 94.
DE RecName: Full=6,7-dimethyl-8-ribityllumazine synthase 2 {ECO:0000255|HAMAP-Rule:MF_00178};
DE Short=DMRL synthase 2 {ECO:0000255|HAMAP-Rule:MF_00178};
DE Short=LS 2 {ECO:0000255|HAMAP-Rule:MF_00178};
DE Short=Lumazine synthase 2 {ECO:0000255|HAMAP-Rule:MF_00178};
DE EC=2.5.1.78 {ECO:0000255|HAMAP-Rule:MF_00178, ECO:0000269|PubMed:10482294, ECO:0000269|PubMed:16165152, ECO:0000269|PubMed:20195542};
DE AltName: Full=BLS {ECO:0000303|PubMed:16455994};
DE AltName: Full=Type II lumazine synthase {ECO:0000303|PubMed:16923880};
GN Name=ribH2 {ECO:0000303|PubMed:16923880, ECO:0000303|PubMed:20195542};
GN Synonyms=ribH-2; OrderedLocusNames=BAB2_0545;
OS Brucella abortus (strain 2308).
OC Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC Brucellaceae; Brucella/Ochrobactrum group; Brucella.
OX NCBI_TaxID=359391;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=2308;
RX PubMed=16299333; DOI=10.1128/iai.73.12.8353-8361.2005;
RA Chain P.S., Comerci D.J., Tolmasky M.E., Larimer F.W., Malfatti S.A.,
RA Vergez L.M., Aguero F., Land M.L., Ugalde R.A., Garcia E.;
RT "Whole-genome analyses of speciation events in pathogenic Brucellae.";
RL Infect. Immun. 73:8353-8361(2005).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOTECHNOLOGY, AND SUBCELLULAR LOCATION.
RX PubMed=10482294; DOI=10.1099/00222615-48-9-833;
RA Goldbaum F.A., Velikovsky C.A., Baldi P.C., Moertl S., Bacher A.,
RA Fossati C.A.;
RT "The 18-kDa cytoplasmic protein of Brucella species -- an antigen useful
RT for diagnosis -- is a lumazine synthase.";
RL J. Med. Microbiol. 48:833-839(1999).
RN [3]
RP FUNCTION, AND BIOTECHNOLOGY.
RX PubMed=14500496; DOI=10.1128/iai.71.10.5750-5755.2003;
RA Velikovsky C.A., Goldbaum F.A., Cassataro J., Estein S., Bowden R.A.,
RA Bruno L., Fossati C.A., Giambartolomei G.H.;
RT "Brucella lumazine synthase elicits a mixed Th1-Th2 immune response and
RT reduces infection in mice challenged with Brucella abortus 544
RT independently of the adjuvant formulation used.";
RL Infect. Immun. 71:5750-5755(2003).
RN [4]
RP SUBUNIT, AND BIOTECHNOLOGY.
RX PubMed=14660615; DOI=10.1074/jbc.m312035200;
RA Zylberman V., Craig P.O., Klinke S., Braden B.C., Cauerhff A.,
RA Goldbaum F.A.;
RT "High order quaternary arrangement confers increased structural stability
RT to Brucella sp. lumazine synthase.";
RL J. Biol. Chem. 279:8093-8101(2004).
RN [5]
RP BIOTECHNOLOGY.
RX PubMed=15390265; DOI=10.1002/prot.20248;
RA Laplagne D.A., Zylberman V., Ainciart N., Steward M.W., Sciutto E.,
RA Fossati C.A., Goldbaum F.A.;
RT "Engineering of a polymeric bacterial protein as a scaffold for the
RT multiple display of peptides.";
RL Proteins 57:820-828(2004).
RN [6]
RP GENE NAME, AND PATHWAY.
RX PubMed=16923880; DOI=10.1128/jb.00207-06;
RA Zylberman V., Klinke S., Haase I., Bacher A., Fischer M., Goldbaum F.A.;
RT "Evolution of vitamin B2 biosynthesis: 6,7-dimethyl-8-ribityllumazine
RT synthases of Brucella.";
RL J. Bacteriol. 188:6135-6142(2006).
RN [7]
RP FUNCTION AS AN IMMUNE MODULATOR, AND BIOTECHNOLOGY.
RX PubMed=16455994; DOI=10.4049/jimmunol.176.4.2366;
RA Berguer P.M., Mundinano J., Piazzon I., Goldbaum F.A.;
RT "A polymeric bacterial protein activates dendritic cells via TLR4.";
RL J. Immunol. 176:2366-2372(2006).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, ROLE IN VIRULENCE, GENE NAME, DISRUPTION
RP PHENOTYPE, MUTAGENESIS OF TRP-20, INDUCTION, AND PATHWAY.
RC STRAIN=2308;
RX PubMed=20195542; DOI=10.1371/journal.pone.0009435;
RA Bonomi H.R., Marchesini M.I., Klinke S., Ugalde J.E., Zylberman V.,
RA Ugalde R.A., Comerci D.J., Goldbaum F.A.;
RT "An atypical riboflavin pathway is essential for Brucella abortus
RT virulence.";
RL PLoS ONE 5:E9435-E9435(2010).
RN [9]
RP FUNCTION AS AN IMMUNE MODULATOR, AND BIOTECHNOLOGY.
RX PubMed=23029192; DOI=10.1371/journal.pone.0045705;
RA Berguer P.M., Alzogaray V.A., Rossi A.H., Mundinano J., Piazzon I.,
RA Goldbaum F.A.;
RT "A polymeric protein induces specific cytotoxicity in a TLR4 dependent
RT manner in the absence of adjuvants.";
RL PLoS ONE 7:E45705-E45705(2012).
RN [10]
RP FUNCTION AS AN IMMUNE MODULATOR, AND BIOTECHNOLOGY.
RX PubMed=25973756; DOI=10.1371/journal.pone.0126827;
RA Rossi A.H., Farias A., Fernandez J.E., Bonomi H.R., Goldbaum F.A.,
RA Berguer P.M.;
RT "Brucella spp. lumazine synthase induces a TLR4-mediated protective
RT response against B16 melanoma in mice.";
RL PLoS ONE 10:E0126827-E0126827(2015).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS) IN COMPLEX WITH PHOSPHATE.
RX PubMed=10764570; DOI=10.1006/jmbi.2000.3640;
RA Braden B.C., Velikovsky C.A., Cauerhff A.A., Polikarpov I., Goldbaum F.A.;
RT "Divergence in macromolecular assembly: X-ray crystallographic structure
RT analysis of lumazine synthase from Brucella abortus.";
RL J. Mol. Biol. 297:1031-1036(2000).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.90 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH
RP SUBSTRATE ANALOG INHIBITOR AND PHOSPHATE, FUNCTION, CATALYTIC ACTIVITY, AND
RP KINETIC PARAMETERS.
RX PubMed=16165152; DOI=10.1016/j.jmb.2005.08.017;
RA Klinke S., Zylberman V., Vega D.R., Guimaraes B.G., Braden B.C.,
RA Goldbaum F.A.;
RT "Crystallographic studies on decameric Brucella spp. lumazine synthase: a
RT novel quaternary arrangement evolved for a new function?";
RL J. Mol. Biol. 353:124-137(2005).
CC -!- FUNCTION: Catalyzes the formation of 6,7-dimethyl-8-ribityllumazine by
CC condensation of 5-amino-6-(D-ribitylamino)uracil with 3,4-dihydroxy-2-
CC butanone 4-phosphate (PubMed:10482294, PubMed:16165152,
CC PubMed:20195542). This is the penultimate step in the biosynthesis of
CC riboflavin. The isozyme RibH2 but not RibH1 is essential for Brucella
CC intracellular survival and replication inside macrophages or in mice
CC (PubMed:20195542). Displays low catalytic activity in comparison with
CC the isozyme RibH1 (PubMed:16165152). Is a highly immunogenic protein
CC (PubMed:14500496). Activates dendritic cells (DCs) in vitro, increasing
CC the levels of costimulatory molecules and the secretion of pro-
CC inflammatory cytokines, and recruits DCs, B cells and CD8+ T cells in
CC vivo, both effects in a TLR4-dependent manner (PubMed:16455994).
CC Induces the cross presentation of covalently attached peptides and
CC generates a strong and long-lasting humoral immune response without
CC adjuvants; TLR4 signaling is necessary for the induction of the
CC cytotoxic response but not for antigen cross presentation
CC (PubMed:23029192). Elicits a TLR4-mediated protective response against
CC B16 melanoma in mice, slowing tumor growth and prolonging mice survival
CC (PubMed:25973756). {ECO:0000269|PubMed:10482294,
CC ECO:0000269|PubMed:14500496, ECO:0000269|PubMed:16165152,
CC ECO:0000269|PubMed:16455994, ECO:0000269|PubMed:20195542,
CC ECO:0000269|PubMed:23029192, ECO:0000269|PubMed:25973756}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2S)-2-hydroxy-3-oxobutyl phosphate + 5-amino-6-(D-
CC ribitylamino)uracil = 6,7-dimethyl-8-(1-D-ribityl)lumazine + H(+) + 2
CC H2O + phosphate; Xref=Rhea:RHEA:26152, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15934, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:58201, ChEBI:CHEBI:58830; EC=2.5.1.78;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00178,
CC ECO:0000269|PubMed:10482294, ECO:0000269|PubMed:16165152,
CC ECO:0000269|PubMed:20195542};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=10 uM for 5-amino-6-(D-ribitylamino)uracil (at 37 degrees Celsius
CC and pH 7.0) {ECO:0000269|PubMed:16165152};
CC KM=4000 uM for 3,4-dihydroxy-2-butanone 4-phosphate (at 37 degrees
CC Celsius and pH 7.0) {ECO:0000269|PubMed:16165152};
CC Vmax=20 nmol/min/mg enzyme (at 37 degrees Celsius and pH 7.0)
CC {ECO:0000269|PubMed:16165152};
CC -!- PATHWAY: Cofactor biosynthesis; riboflavin biosynthesis; riboflavin
CC from 2-hydroxy-3-oxobutyl phosphate and 5-amino-6-(D-
CC ribitylamino)uracil: step 1/2. {ECO:0000255|HAMAP-Rule:MF_00178,
CC ECO:0000305|PubMed:16923880, ECO:0000305|PubMed:20195542}.
CC -!- SUBUNIT: Homodecamer, arranged as a dimer of pentamers.
CC {ECO:0000269|PubMed:10764570, ECO:0000269|PubMed:14660615,
CC ECO:0000269|PubMed:16165152}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:10482294}.
CC -!- INDUCTION: The two ribH genes may be differentially expressed during
CC the Brucella infection cycle. Brucella would use RibH1 for flavin
CC biosynthesis during the extracellular phase and RibH2 during
CC intracellular growth. {ECO:0000305|PubMed:20195542}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene are not auxotrophic for
CC riboflavin and grow at wild-type rates in both rich and minimal media.
CC But simultaneous disruption of ribH1 and ribH2 is lethal. The ribH2
CC mutant shows a decrease in survival and replication in macrophages at
CC all time-points and is attenuated in mice.
CC {ECO:0000269|PubMed:20195542}.
CC -!- BIOTECHNOLOGY: Could be useful as a specific antigen for the
CC serological diagnosis of active infection of both human and bovine
CC brucellosis (PubMed:10482294). Has been used as a protein carrier of
CC foreign peptides and proteins (PubMed:15390265). The described
CC characteristics of BLS make this protein an ideal antigen carrier for
CC vaccine development (PubMed:14660615, PubMed:15390265, PubMed:16455994,
CC PubMed:23029192, PubMed:14500496). The antitumor effect of BLS could
CC lead to a therapeutic strategy utilizing a TLR4 ligand; as the
CC expression of TLR4 has been reported on a large number of tumors, BLS
CC signaling via TLR4 could make a notable contribution to the success of
CC cancer treatment when coadministered with other cancer vaccines or
CC treatments like radiation or chemotherapy (PubMed:25973756).
CC {ECO:0000269|PubMed:10482294, ECO:0000269|PubMed:14500496,
CC ECO:0000269|PubMed:14660615, ECO:0000269|PubMed:15390265,
CC ECO:0000269|PubMed:16455994, ECO:0000269|PubMed:23029192,
CC ECO:0000269|PubMed:25973756}.
CC -!- SIMILARITY: Belongs to the DMRL synthase family. {ECO:0000255|HAMAP-
CC Rule:MF_00178}.
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DR EMBL; AM040265; CAJ12711.1; -; Genomic_DNA.
DR RefSeq; WP_002965946.1; NZ_KN046823.1.
DR PDB; 1DI0; X-ray; 2.70 A; A/B/C/D/E=1-158.
DR PDB; 1T13; X-ray; 2.90 A; A/B/C/D/E=1-158.
DR PDB; 1XN1; X-ray; 3.05 A; A/B/C/D/E/F/G/H/I/J=1-158.
DR PDBsum; 1DI0; -.
DR PDBsum; 1T13; -.
DR PDBsum; 1XN1; -.
DR AlphaFoldDB; Q2YKV1; -.
DR SMR; Q2YKV1; -.
DR STRING; 359391.BAB2_0545; -.
DR EnsemblBacteria; CAJ12711; CAJ12711; BAB2_0545.
DR GeneID; 45126031; -.
DR GeneID; 55592351; -.
DR KEGG; bmf:BAB2_0545; -.
DR PATRIC; fig|359391.11.peg.2737; -.
DR HOGENOM; CLU_089358_0_0_5; -.
DR OMA; PHHFHEH; -.
DR PhylomeDB; Q2YKV1; -.
DR BRENDA; 2.5.1.78; 994.
DR UniPathway; UPA00275; UER00404.
DR Proteomes; UP000002719; Chromosome II.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0009349; C:riboflavin synthase complex; IEA:InterPro.
DR GO; GO:0000906; F:6,7-dimethyl-8-ribityllumazine synthase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0009231; P:riboflavin biosynthetic process; IEA:UniProtKB-UniRule.
DR Gene3D; 3.40.50.960; -; 1.
DR HAMAP; MF_00178; Lumazine_synth; 1.
DR InterPro; IPR034964; LS.
DR InterPro; IPR002180; LS/RS.
DR InterPro; IPR036467; LS/RS_sf.
DR PANTHER; PTHR21058; PTHR21058; 1.
DR Pfam; PF00885; DMRL_synthase; 1.
DR SUPFAM; SSF52121; SSF52121; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Reference proteome; Riboflavin biosynthesis;
KW Transferase; Virulence.
FT CHAIN 1..158
FT /note="6,7-dimethyl-8-ribityllumazine synthase 2"
FT /id="PRO_0000425958"
FT ACT_SITE 86
FT /note="Proton donor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00178"
FT BINDING 20
FT /ligand="5-amino-6-(D-ribitylamino)uracil"
FT /ligand_id="ChEBI:CHEBI:15934"
FT /evidence="ECO:0000305|PubMed:16165152"
FT BINDING 54..56
FT /ligand="5-amino-6-(D-ribitylamino)uracil"
FT /ligand_id="ChEBI:CHEBI:15934"
FT /evidence="ECO:0000305|PubMed:16165152"
FT BINDING 78..80
FT /ligand="5-amino-6-(D-ribitylamino)uracil"
FT /ligand_id="ChEBI:CHEBI:15934"
FT /evidence="ECO:0000305|PubMed:16165152"
FT BINDING 111
FT /ligand="5-amino-6-(D-ribitylamino)uracil"
FT /ligand_id="ChEBI:CHEBI:15934"
FT /evidence="ECO:0000305|PubMed:16165152"
FT BINDING 125
FT /ligand="(2S)-2-hydroxy-3-oxobutyl phosphate"
FT /ligand_id="ChEBI:CHEBI:58830"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00178"
FT MUTAGEN 20
FT /note="W->A: Loss of enzymatic activity."
FT /evidence="ECO:0000269|PubMed:20195542"
FT STRAND 11..18
FT /evidence="ECO:0007829|PDB:1DI0"
FT HELIX 22..40
FT /evidence="ECO:0007829|PDB:1DI0"
FT STRAND 43..53
FT /evidence="ECO:0007829|PDB:1DI0"
FT HELIX 54..56
FT /evidence="ECO:0007829|PDB:1DI0"
FT HELIX 57..66
FT /evidence="ECO:0007829|PDB:1DI0"
FT STRAND 71..78
FT /evidence="ECO:0007829|PDB:1DI0"
FT STRAND 83..85
FT /evidence="ECO:0007829|PDB:1DI0"
FT HELIX 88..105
FT /evidence="ECO:0007829|PDB:1DI0"
FT STRAND 109..114
FT /evidence="ECO:0007829|PDB:1DI0"
FT STRAND 116..118
FT /evidence="ECO:0007829|PDB:1DI0"
FT HELIX 123..153
FT /evidence="ECO:0007829|PDB:1DI0"
SQ SEQUENCE 158 AA; 17356 MW; EE59C2C815E53A2B CRC64;
MNQSCPNKTS FKIAFIQARW HADIVDEARK SFVAELAAKT GGSVEVEIFD VPGAYEIPLH
AKTLARTGRY AAIVGAAFVI DGGIYRHDFV ATAVINGMMQ VQLETEVPVL SVVLTPHHFH
ESKEHHDFFH AHFKVKGVEA AHAALQIVSE RSRIAALV
