ID   RMLA_SALTY              Reviewed;         292 AA.
AC   P26393;
DT   01-AUG-1992, integrated into UniProtKB/Swiss-Prot.
DT   01-AUG-1992, sequence version 1.
DT   27-MAR-2024, entry version 143.
DE   RecName: Full=Glucose-1-phosphate thymidylyltransferase {ECO:0000303|PubMed:8382158};
DE            EC=2.7.7.24 {ECO:0000269|PubMed:8382158};
DE   AltName: Full=dTDP-glucose pyrophosphorylase;
DE            Short=Ep;
DE   AltName: Full=dTDP-glucose synthase;
GN   Name=rmlA; Synonyms=rfbA {ECO:0000303|PubMed:8382158};
GN   OrderedLocusNames=STM2095;
OS   Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720).
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Salmonella.
OX   NCBI_TaxID=99287;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=LT2;
RX   PubMed=1710759; DOI=10.1111/j.1365-2958.1991.tb00741.x;
RA   Jiang X.-M., Neal B., Santiago F., Lee S.J., Romana L.K., Reeves P.R.;
RT   "Structure and sequence of the rfb (O antigen) gene cluster of Salmonella
RT   serovar typhimurium (strain LT2).";
RL   Mol. Microbiol. 5:695-713(1991).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=LT2 / SGSC1412 / ATCC 700720;
RX   PubMed=11677609; DOI=10.1038/35101614;
RA   McClelland M., Sanderson K.E., Spieth J., Clifton S.W., Latreille P.,
RA   Courtney L., Porwollik S., Ali J., Dante M., Du F., Hou S., Layman D.,
RA   Leonard S., Nguyen C., Scott K., Holmes A., Grewal N., Mulvaney E.,
RA   Ryan E., Sun H., Florea L., Miller W., Stoneking T., Nhan M., Waterston R.,
RA   Wilson R.K.;
RT   "Complete genome sequence of Salmonella enterica serovar Typhimurium LT2.";
RL   Nature 413:852-856(2001).
RN   [3]
RP   PROTEIN SEQUENCE OF 1-22, FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE
RP   SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RC   STRAIN=LT2;
RX   PubMed=8382158; DOI=10.1111/j.1432-1033.1993.tb17607.x;
RA   Lindquist L., Kaiser R., Reeves P.R., Lindberg A.A.;
RT   "Purification, characterization and HPLC assay of Salmonella glucose-1-
RT   phosphate thymidylyltransferase from the cloned rfbA gene.";
RL   Eur. J. Biochem. 211:763-770(1993).
RN   [4]
RP   CHARACTERIZATION, MUTAGENESIS OF THR-201 AND TRP-224, AND X-RAY
RP   CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF COMPLEXES WITH UDP-GLC AND DTTP.
RC   STRAIN=LT2;
RX   PubMed=11373625; DOI=10.1038/88618;
RA   Barton W.A., Lesniak J., Biggins J.B., Jeffrey P.D., Jiang J.,
RA   Rajashankar K.R., Thorson J.S., Nikolov D.B.;
RT   "Structure, mechanism and engineering of a nucleotidylyltransferase as a
RT   first step toward glycorandomization.";
RL   Nat. Struct. Biol. 8:545-551(2001).
CC   -!- FUNCTION: Catalyzes the formation of dTDP-glucose, from dTTP and
CC       glucose 1-phosphate, as well as its pyrophosphorolysis. Is the first of
CC       four enzymes commited to biosynthesis of dTDP-L-rhamnose in
CC       S.typhimurium LT2. Is also able to convert non natural substrates such
CC       as a wide array of alpha-D-hexopyranosyl, deoxy-alpha-D-glucopyranosyl,
CC       aminodeoxy-alpha-D-hexopyranosyl and acetamidodeoxy-alpha-D-
CC       hexopyranosyl phosphates to their corresponding dTDP- and UDP-
CC       nucleotide sugars. {ECO:0000269|PubMed:8382158}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=alpha-D-glucose 1-phosphate + dTTP + H(+) = diphosphate +
CC         dTDP-alpha-D-glucose; Xref=Rhea:RHEA:15225, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:33019, ChEBI:CHEBI:37568, ChEBI:CHEBI:57477,
CC         ChEBI:CHEBI:58601; EC=2.7.7.24;
CC         Evidence={ECO:0000269|PubMed:8382158};
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000305};
CC       Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000305};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=0.02 mM for dTTP {ECO:0000269|PubMed:8382158};
CC         KM=0.11 mM for alpha-D-glucose 1-phosphate
CC         {ECO:0000269|PubMed:8382158};
CC         KM=0.083 mM for dTDP-alpha-D-glucose {ECO:0000269|PubMed:8382158};
CC         KM=0.15 mM for diphosphate {ECO:0000269|PubMed:8382158};
CC         KM=0.7 mM for dTTP;
CC         KM=0.3 mM for G1P;
CC         Vmax=54.7 umol/min/mg enzyme for the formation of
CC         dTDP-alpha-D-glucose {ECO:0000269|PubMed:8382158};
CC         Vmax=329 umol/min/mg enzyme for the pyrophosphorolysis of
CC         dTDP-alpha-D-glucose {ECO:0000269|PubMed:8382158};
CC   -!- PATHWAY: Carbohydrate biosynthesis; dTDP-L-rhamnose biosynthesis.
CC       {ECO:0000305|PubMed:8382158}.
CC   -!- PATHWAY: Bacterial outer membrane biogenesis; LPS O-antigen
CC       biosynthesis.
CC   -!- SUBUNIT: Homotetramer.
CC   -!- SIMILARITY: Belongs to the glucose-1-phosphate thymidylyltransferase
CC       family. {ECO:0000305}.
CC   -!- CAUTION: The position of the magnesium ion observed in the
CC       crystallographic structure is different from that observed in the
CC       structure of E.coli RmlA2 (RffH) and does not seem to correspond to the
CC       binding site for the catalytically essential magnesium ion. Therefore,
CC       we choose to propagate in the feature lines the positions found in the
CC       E.coli structure. {ECO:0000305}.
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; X56793; CAA40117.1; -; Genomic_DNA.
DR   EMBL; AE006468; AAL20999.1; -; Genomic_DNA.
DR   PIR; S15301; S15301.
DR   RefSeq; NP_461040.1; NC_003197.2.
DR   RefSeq; WP_000857529.1; NC_003197.2.
DR   PDB; 1IIM; X-ray; 2.10 A; A/B=1-292.
DR   PDB; 1IIN; X-ray; 2.10 A; A/B/C/D=1-292.
DR   PDB; 1MP3; X-ray; 2.20 A; A/B=1-292.
DR   PDB; 1MP4; X-ray; 2.20 A; A/B=1-292.
DR   PDB; 1MP5; X-ray; 2.75 A; A/B/C/D=1-292.
DR   PDB; 3PKP; X-ray; 2.60 A; A/B/C/D/I/J/K/L=1-292.
DR   PDB; 3PKQ; X-ray; 2.40 A; A/B/C/D=1-292.
DR   PDBsum; 1IIM; -.
DR   PDBsum; 1IIN; -.
DR   PDBsum; 1MP3; -.
DR   PDBsum; 1MP4; -.
DR   PDBsum; 1MP5; -.
DR   PDBsum; 3PKP; -.
DR   PDBsum; 3PKQ; -.
DR   AlphaFoldDB; P26393; -.
DR   SMR; P26393; -.
DR   STRING; 99287.STM2095; -.
DR   DrugBank; DB02452; Thymidine 5'-triphosphate.
DR   DrugBank; DB01861; Uridine diphosphate glucose.
DR   PaxDb; 99287-STM2095; -.
DR   GeneID; 1253616; -.
DR   KEGG; stm:STM2095; -.
DR   PATRIC; fig|99287.12.peg.2217; -.
DR   HOGENOM; CLU_029499_9_0_6; -.
DR   OMA; PFIMYLG; -.
DR   PhylomeDB; P26393; -.
DR   BioCyc; SENT99287:STM2095-MONOMER; -.
DR   SABIO-RK; P26393; -.
DR   UniPathway; UPA00124; -.
DR   UniPathway; UPA00281; -.
DR   EvolutionaryTrace; P26393; -.
DR   Proteomes; UP000001014; Chromosome.
DR   GO; GO:0008879; F:glucose-1-phosphate thymidylyltransferase activity; IDA:UniProtKB.
DR   GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR   GO; GO:0019305; P:dTDP-rhamnose biosynthetic process; IEA:UniProtKB-UniPathway.
DR   GO; GO:0045226; P:extracellular polysaccharide biosynthetic process; IDA:UniProtKB.
DR   GO; GO:0009243; P:O antigen biosynthetic process; IEA:UniProtKB-UniPathway.
DR   CDD; cd02538; G1P_TT_short; 1.
DR   InterPro; IPR005907; G1P_thy_trans_s.
DR   InterPro; IPR005835; NTP_transferase_dom.
DR   InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR   NCBIfam; TIGR01207; rmlA; 1.
DR   PANTHER; PTHR43532; GLUCOSE-1-PHOSPHATE THYMIDYLYLTRANSFERASE; 1.
DR   PANTHER; PTHR43532:SF1; GLUCOSE-1-PHOSPHATE THYMIDYLYLTRANSFERASE 1; 1.
DR   Pfam; PF00483; NTP_transferase; 1.
DR   SUPFAM; SSF53448; Nucleotide-diphospho-sugar transferases; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Direct protein sequencing; Lipopolysaccharide biosynthesis;
KW   Magnesium; Metal-binding; Nucleotidyltransferase; Reference proteome;
KW   Transferase.
FT   CHAIN           1..292
FT                   /note="Glucose-1-phosphate thymidylyltransferase"
FT                   /id="PRO_0000207995"
FT   BINDING         111
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /evidence="ECO:0000250|UniProtKB:P61887"
FT   BINDING         226
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT                   /evidence="ECO:0000250|UniProtKB:P61887"
FT   MUTAGEN         201
FT                   /note="T->A: Two-fold increase in the conversion of
FT                   2-acetamido-2-deoxy-alpha-D-glucopyranosyl phosphate."
FT                   /evidence="ECO:0000269|PubMed:11373625"
FT   MUTAGEN         224
FT                   /note="W->H: Is able to convert both
FT                   6-acetamido-6-deoxy-alpha-D-glucopyranosyl phosphate and
FT                   alpha-D-glucopyranuronic acid 1-(dihydrogen phosphate),
FT                   which are not accepted by the wild-type."
FT                   /evidence="ECO:0000269|PubMed:11373625"
FT   STRAND          5..9
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           15..17
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           20..22
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           26..28
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          29..31
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           38..46
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          51..56
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   TURN            58..60
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           61..68
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           72..74
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          77..82
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          84..87
FT                   /evidence="ECO:0007829|PDB:3PKP"
FT   HELIX           89..91
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           92..95
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           97..100
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          105..109
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          112..115
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           119..128
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          131..139
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           143..145
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          146..151
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          153..155
FT                   /evidence="ECO:0007829|PDB:1IIN"
FT   STRAND          157..163
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          168..179
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           183..189
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           200..209
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          213..217
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   STRAND          222..226
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           230..247
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           254..260
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           266..273
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           274..276
FT                   /evidence="ECO:0007829|PDB:1IIM"
FT   HELIX           280..288
FT                   /evidence="ECO:0007829|PDB:1IIM"
SQ   SEQUENCE   292 AA;  32453 MW;  11065BA9DF2B0268 CRC64;
     MKTRKGIILA GGSGTRLYPV TMAVSKQLLP IYDKPMIYYP LSTLMLAGIR DILIISTPQD
     TPRFQQLLGD GSQWGLNLQY KVQPSPDGLA QAFIIGEEFI GHDDCALVLG DNIFYGHDLP
     KLMEAAVNKE SGATVFAYHV NDPERYGVVE FDQKGTAVSL EEKPLQPKSN YAVTGLYFYD
     NSVVEMAKNL KPSARGELEI TDINRIYMEQ GRLSVAMMGR GYAWLDTGTH QSLIEASNFI
     ATIEERQGLK VSCPEEIAFR KNFINAQQVI ELAGPLSKND YGKYLLKMVK GL
//