RMLA_SALTY
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 03-AUG-2022, entry version 135.
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; Proteobacteria; 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=0.020 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 dTDP-alpha-D-
CC glucose {ECO:0000269|PubMed:8382158};
CC Vmax=329 umol/min/mg enzyme for the pyrophosphorolysis of dTDP-alpha-
CC 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}.
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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; P26393; -.
DR EnsemblBacteria; AAL20999; AAL20999; 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-MON; -.
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 Gene3D; 3.90.550.10; -; 1.
DR InterPro; IPR005907; G1P_thy_trans_s.
DR InterPro; IPR005835; NTP_transferase_dom.
DR InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR PANTHER; PTHR43532; PTHR43532; 1.
DR Pfam; PF00483; NTP_transferase; 1.
DR SUPFAM; SSF53448; SSF53448; 1.
DR TIGRFAMs; TIGR01207; rmlA; 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"
FT BINDING 226
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT MUTAGEN 201
FT /note="T->A: Two-fold increase in the conversion of 2-
FT 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 6-acetamido-6-deoxy-
FT alpha-D-glucopyranosyl phosphate and alpha-D-
FT glucopyranuronic acid 1-(dihydrogen phosphate), which are
FT 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
