THIE_BACSU
ID THIE_BACSU Reviewed; 222 AA.
AC P39594;
DT 01-FEB-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1995, sequence version 1.
DT 03-AUG-2022, entry version 151.
DE RecName: Full=Thiamine-phosphate synthase;
DE Short=TP synthase;
DE Short=TPS;
DE EC=2.5.1.3 {ECO:0000269|PubMed:9139923};
DE AltName: Full=Thiamine-phosphate pyrophosphorylase;
DE Short=TMP pyrophosphorylase;
DE Short=TMP-PPase;
GN Name=thiE; Synonyms=thiC, ywbK; OrderedLocusNames=BSU38290;
GN ORFNames=ipa-26d;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=7934828; DOI=10.1111/j.1365-2958.1993.tb01963.x;
RA Glaser P., Kunst F., Arnaud M., Coudart M.P., Gonzales W., Hullo M.-F.,
RA Ionescu M., Lubochinsky B., Marcelino L., Moszer I., Presecan E.,
RA Santana M., Schneider E., Schweizer J., Vertes A., Rapoport G., Danchin A.;
RT "Bacillus subtilis genome project: cloning and sequencing of the 97 kb
RT region from 325 degrees to 333 degrees.";
RL Mol. Microbiol. 10:371-384(1993).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, KINETIC PARAMETERS, AND INDUCTION.
RC STRAIN=168 / CU1065;
RX PubMed=9139923; DOI=10.1128/jb.179.9.3030-3035.1997;
RA Zhang Y., Taylor S.V., Chiu H.-J., Begley T.P.;
RT "Characterization of the Bacillus subtilis thiC operon involved in thiamine
RT biosynthesis.";
RL J. Bacteriol. 179:3030-3035(1997).
RN [4]
RP FUNCTION, SUBSTRATE SPECIFICITY, MUTAGENESIS OF SER-117, AND REACTION
RP MECHANISM.
RC STRAIN=168 / CU1065;
RX PubMed=11513588; DOI=10.1021/bi010267q;
RA Reddick J.J., Nicewonger R., Begley T.P.;
RT "Mechanistic studies on thiamin phosphate synthase: evidence for a
RT dissociative mechanism.";
RL Biochemistry 40:10095-10102(2001).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.25 ANGSTROMS) IN COMPLEX WITH THIAMINE PHOSPHATE
RP AND MAGNESIUM PYROPHOSPHATE.
RC STRAIN=168 / CU1065;
RX PubMed=10350464; DOI=10.1021/bi982903z;
RA Chiu H.-J., Reddick J.J., Begley T.P., Ealick S.E.;
RT "Crystal structure of thiamin phosphate synthase from Bacillus subtilis at
RT 1.25 A resolution.";
RL Biochemistry 38:6460-6470(1999).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS) OF WILD-TYPE AND MUTANT ALA-117 IN
RP COMPLEXES WITH SUBSTRATE; PRODUCTS AND INTERMEDIATES.
RC STRAIN=168 / CU1065;
RX PubMed=11513589; DOI=10.1021/bi0104726;
RA Peapus D.H., Chiu H.J., Campobasso N., Reddick J.J., Begley T.P.,
RA Ealick S.E.;
RT "Structural characterization of the enzyme-substrate, enzyme-intermediate,
RT and enzyme-product complexes of thiamin phosphate synthase.";
RL Biochemistry 40:10103-10114(2001).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS).
RA McCulloch K.M., Hanes J.W., Abdelwahed S., Mahanta N., Hazra A., Ishida K.,
RA Begley T.P., Ealick S.E.;
RT "Crystal structure and kinetic characterization of Bacillus subtilis
RT thiamin phosphate synthase with a carboxylated thiazole phosphate.";
RL Submitted (JUL-2011) to the PDB data bank.
CC -!- FUNCTION: Condenses 4-methyl-5-(beta-hydroxyethyl)thiazole
CC monophosphate (THZ-P) and 2-methyl-4-amino-5-hydroxymethyl pyrimidine
CC pyrophosphate (HMP-PP) to form thiamine monophosphate (TMP). Is also
CC able to use the 2-methoxy analog MeO-HMP-PP, as substrate in vitro, but
CC not the 2-trifluoromethyl analog CF(3)-HMP-PP.
CC {ECO:0000269|PubMed:11513588, ECO:0000269|PubMed:9139923}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-ylidene]ethyl
CC phosphate + 4-amino-2-methyl-5-(diphosphooxymethyl)pyrimidine + 2
CC H(+) = CO2 + diphosphate + thiamine phosphate; Xref=Rhea:RHEA:47844,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:16526, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:37575, ChEBI:CHEBI:57841, ChEBI:CHEBI:62899; EC=2.5.1.3;
CC Evidence={ECO:0000269|PubMed:9139923};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-(2-carboxy-4-methylthiazol-5-yl)ethyl phosphate + 4-amino-2-
CC methyl-5-(diphosphooxymethyl)pyrimidine + 2 H(+) = CO2 + diphosphate
CC + thiamine phosphate; Xref=Rhea:RHEA:47848, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:33019, ChEBI:CHEBI:37575,
CC ChEBI:CHEBI:57841, ChEBI:CHEBI:62890; EC=2.5.1.3;
CC Evidence={ECO:0000269|PubMed:9139923};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-amino-2-methyl-5-(diphosphooxymethyl)pyrimidine + 4-methyl-
CC 5-(2-phosphooxyethyl)-thiazole + H(+) = diphosphate + thiamine
CC phosphate; Xref=Rhea:RHEA:22328, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:37575, ChEBI:CHEBI:57841,
CC ChEBI:CHEBI:58296; EC=2.5.1.3; Evidence={ECO:0000269|PubMed:9139923};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:9139923};
CC Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000269|PubMed:9139923};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.7 uM for HMP-PP {ECO:0000269|PubMed:9139923};
CC KM=1.2 uM for THZ-P {ECO:0000269|PubMed:9139923};
CC Vmax=0.7 umol/min/mg enzyme {ECO:0000269|PubMed:9139923};
CC -!- PATHWAY: Cofactor biosynthesis; thiamine diphosphate biosynthesis;
CC thiamine phosphate from 4-amino-2-methyl-5-diphosphomethylpyrimidine
CC and 4-methyl-5-(2-phosphoethyl)-thiazole: step 1/1.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:10350464}.
CC -!- INDUCTION: Is weakly repressed by THZ and not at all by thiamine.
CC {ECO:0000269|PubMed:9139923}.
CC -!- MISCELLANEOUS: Thiamine phosphate synthase appears to proceed with a
CC dissociative mechanism (SN1 like) in which the pyrimidine pyrophosphate
CC dissociates to give a reactive pyrimidine intermediate which is then
CC trapped by the thiazole moiety.
CC -!- SIMILARITY: Belongs to the thiamine-phosphate synthase family.
CC {ECO:0000305}.
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DR EMBL; X73124; CAA51582.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB15855.1; -; Genomic_DNA.
DR PIR; S39681; S39681.
DR RefSeq; NP_391708.1; NC_000964.3.
DR RefSeq; WP_003244128.1; NZ_JNCM01000034.1.
DR PDB; 1G4E; X-ray; 1.60 A; A/B=1-222.
DR PDB; 1G4P; X-ray; 2.50 A; A/B=1-222.
DR PDB; 1G4S; X-ray; 1.70 A; A/B=1-222.
DR PDB; 1G4T; X-ray; 1.55 A; A/B=1-222.
DR PDB; 1G67; X-ray; 1.40 A; A/B=1-222.
DR PDB; 1G69; X-ray; 1.50 A; A/B=1-222.
DR PDB; 1G6C; X-ray; 1.40 A; A/B=1-222.
DR PDB; 2TPS; X-ray; 1.25 A; A/B=1-222.
DR PDB; 3O15; X-ray; 1.95 A; A=1-222.
DR PDB; 3O16; X-ray; 2.10 A; A=1-222.
DR PDBsum; 1G4E; -.
DR PDBsum; 1G4P; -.
DR PDBsum; 1G4S; -.
DR PDBsum; 1G4T; -.
DR PDBsum; 1G67; -.
DR PDBsum; 1G69; -.
DR PDBsum; 1G6C; -.
DR PDBsum; 2TPS; -.
DR PDBsum; 3O15; -.
DR PDBsum; 3O16; -.
DR AlphaFoldDB; P39594; -.
DR SMR; P39594; -.
DR STRING; 224308.BSU38290; -.
DR DrugBank; DB07782; 4-AMINO-2-TRIFLUOROMETHYL-5-HYDROXYMETHYLPYRIMIDINE PYROPHOSPHATE.
DR DrugBank; DB01788; 4-Imino-5-Methidyl-2-Methylpyrimidine.
DR DrugBank; DB02885; 4-Imino-5-Methidyl-2-Trifluoromethylpyrimidine.
DR DrugBank; DB03145; 4-Methyl-5-Hydroxyethylthiazole Phosphate.
DR DrugBank; DB03416; Thiamine(1+) monophosphate.
DR DrugBank; DB02254; Trifluoro-thiamin phosphate.
DR PaxDb; P39594; -.
DR PRIDE; P39594; -.
DR EnsemblBacteria; CAB15855; CAB15855; BSU_38290.
DR GeneID; 937316; -.
DR KEGG; bsu:BSU38290; -.
DR PATRIC; fig|224308.179.peg.4145; -.
DR eggNOG; COG0352; Bacteria.
DR InParanoid; P39594; -.
DR OMA; ITAFQFR; -.
DR PhylomeDB; P39594; -.
DR BioCyc; BSUB:BSU38290-MON; -.
DR BioCyc; MetaCyc:BSU38290-MON; -.
DR BRENDA; 2.5.1.3; 658.
DR UniPathway; UPA00060; UER00141.
DR EvolutionaryTrace; P39594; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central.
DR GO; GO:0000287; F:magnesium ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0004789; F:thiamine-phosphate diphosphorylase activity; IBA:GO_Central.
DR GO; GO:0009228; P:thiamine biosynthetic process; IBA:GO_Central.
DR GO; GO:0009229; P:thiamine diphosphate biosynthetic process; IEA:UniProtKB-UniPathway.
DR CDD; cd00564; TMP_TenI; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR HAMAP; MF_00097; TMP_synthase; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR036206; ThiamineP_synth_sf.
DR InterPro; IPR022998; ThiamineP_synth_TenI.
DR InterPro; IPR034291; TMP_synthase.
DR Pfam; PF02581; TMP-TENI; 1.
DR SUPFAM; SSF51391; SSF51391; 1.
DR TIGRFAMs; TIGR00693; thiE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Magnesium; Metal-binding; Reference proteome;
KW Thiamine biosynthesis; Transferase.
FT CHAIN 1..222
FT /note="Thiamine-phosphate synthase"
FT /id="PRO_0000156995"
FT BINDING 44..48
FT /ligand="4-amino-2-methyl-5-(diphosphooxymethyl)pyrimidine"
FT /ligand_id="ChEBI:CHEBI:57841"
FT BINDING 79
FT /ligand="4-amino-2-methyl-5-(diphosphooxymethyl)pyrimidine"
FT /ligand_id="ChEBI:CHEBI:57841"
FT BINDING 80
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT BINDING 99
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT BINDING 117
FT /ligand="4-amino-2-methyl-5-(diphosphooxymethyl)pyrimidine"
FT /ligand_id="ChEBI:CHEBI:57841"
FT BINDING 143..145
FT /ligand="2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-
FT ylidene]ethyl phosphate"
FT /ligand_id="ChEBI:CHEBI:62899"
FT BINDING 146
FT /ligand="4-amino-2-methyl-5-(diphosphooxymethyl)pyrimidine"
FT /ligand_id="ChEBI:CHEBI:57841"
FT BINDING 175
FT /ligand="2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-
FT ylidene]ethyl phosphate"
FT /ligand_id="ChEBI:CHEBI:62899"
FT BINDING 195..196
FT /ligand="2-[(2R,5Z)-2-carboxy-4-methylthiazol-5(2H)-
FT ylidene]ethyl phosphate"
FT /ligand_id="ChEBI:CHEBI:62899"
FT MUTAGEN 117
FT /note="S->A: 8000-fold reduction in catalytic activity."
FT /evidence="ECO:0000269|PubMed:11513588"
FT HELIX 6..12
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 15..19
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 21..23
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 28..38
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 41..45
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 55..72
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 76..80
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 82..88
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 91..95
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 97..99
FT /evidence="ECO:0007829|PDB:1G69"
FT HELIX 102..109
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 112..118
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 121..130
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 133..137
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 145..148
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 156..163
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 170..175
FT /evidence="ECO:0007829|PDB:2TPS"
FT TURN 178..180
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 182..186
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 190..195
FT /evidence="ECO:0007829|PDB:2TPS"
FT HELIX 196..199
FT /evidence="ECO:0007829|PDB:2TPS"
FT STRAND 201..203
FT /evidence="ECO:0007829|PDB:3O16"
FT HELIX 204..221
FT /evidence="ECO:0007829|PDB:2TPS"
SQ SEQUENCE 222 AA; 23681 MW; BF09EB73866F4FB4 CRC64;
MTRISREMMK ELLSVYFIMG SNNTKADPVT VVQKALKGGA TLYQFREKGG DALTGEARIK
FAEKAQAACR EAGVPFIVND DVELALNLKA DGIHIGQEDA NAKEVRAAIG DMILGVSAHT
MSEVKQAEED GADYVGLGPI YPTETKKDTR AVQGVSLIEA VRRQGISIPI VGIGGITIDN
AAPVIQAGAD GVSMISAISQ AEDPESAARK FREEIQTYKT GR
