RMLA1_ECOLI
ID RMLA1_ECOLI Reviewed; 293 AA.
AC P37744; P78081;
DT 01-OCT-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1997, sequence version 2.
DT 03-AUG-2022, entry version 165.
DE RecName: Full=Glucose-1-phosphate thymidylyltransferase 1;
DE Short=G1P-TT 1;
DE EC=2.7.7.24 {ECO:0000269|PubMed:11697907};
DE AltName: Full=dTDP-glucose pyrophosphorylase 1;
DE AltName: Full=dTDP-glucose synthase 1;
GN Name=rfbA; Synonyms=rmlA, rmlA1; OrderedLocusNames=b2039, JW2024;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12 / WG1;
RX PubMed=7517391; DOI=10.1128/jb.176.13.4144-4156.1994;
RA Stevenson G., Neal B., Liu D., Hobbs M., Packer N.H., Batley M.,
RA Redmond J.W., Lindquist L., Reeves P.R.;
RT "Structure of the O antigen of Escherichia coli K-12 and the sequence of
RT its rfb gene cluster.";
RL J. Bacteriol. 176:4144-4156(1994).
RN [2]
RP SEQUENCE REVISION TO 288.
RA Stevenson G.;
RL Submitted (DEC-1997) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=9097040; DOI=10.1093/dnares/3.6.379;
RA Itoh T., Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K.,
RA Kasai H., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T.,
RA Mizobuchi K., Mori H., Mori T., Motomura K., Nakade S., Nakamura Y.,
RA Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y.,
RA Sivasundaram S., Tagami H., Takeda J., Takemoto K., Wada C., Yamamoto Y.,
RA Horiuchi T.;
RT "A 460-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 40.1-50.0 min region on the linkage map.";
RL DNA Res. 3:379-392(1996).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [6]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 247-293.
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=7517390; DOI=10.1128/jb.176.13.4133-4143.1994;
RA Yao Z., Valvano M.A.;
RT "Genetic analysis of the O-specific lipopolysaccharide biosynthesis region
RT (rfb) of Escherichia coli K-12 W3110: identification of genes that confer
RT group 6 specificity to Shigella flexneri serotypes Y and 4a.";
RL J. Bacteriol. 176:4133-4143(1994).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP PATHWAY, SUBUNIT, AND X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF COMPLEXES
RP WITH DTDP-G; DTMP AND G1P AND DEOXYTHYMIDINE AND G1P.
RC STRAIN=K12;
RX PubMed=11697907; DOI=10.1006/jmbi.2001.5073;
RA Zuccotti S., Zanardi D., Rosano C., Sturla L., Tonetti M., Bolognesi M.;
RT "Kinetic and crystallographic analyses support a sequential-ordered bi bi
RT catalytic mechanism for Escherichia coli glucose-1-phosphate
RT thymidylyltransferase.";
RL J. Mol. Biol. 313:831-843(2001).
CC -!- FUNCTION: Catalyzes the formation of dTDP-glucose, from dTTP and
CC glucose 1-phosphate, as well as its pyrophosphorolysis.
CC {ECO:0000269|PubMed:11697907}.
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:11697907};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:11697907};
CC Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000269|PubMed:11697907};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=20.5 uM for dTTP {ECO:0000269|PubMed:11697907};
CC KM=34 uM for glucose 1-phosphate {ECO:0000269|PubMed:11697907};
CC KM=95 uM for dTDP-glucose {ECO:0000269|PubMed:11697907};
CC KM=154 uM for PPi {ECO:0000269|PubMed:11697907};
CC Vmax=194 umol/min/mg enzyme for the forward reaction
CC {ECO:0000269|PubMed:11697907};
CC Vmax=360 umol/min/mg enzyme for the reverse reaction
CC {ECO:0000269|PubMed:11697907};
CC pH dependence:
CC Optimum pH is 8.0-8.5. Active from pH 6.0 to 10.0.
CC {ECO:0000269|PubMed:11697907};
CC -!- PATHWAY: Carbohydrate biosynthesis; dTDP-L-rhamnose biosynthesis.
CC {ECO:0000269|PubMed:11697907}.
CC -!- PATHWAY: Bacterial outer membrane biogenesis; LPS O-antigen
CC biosynthesis. {ECO:0000269|PubMed:11697907}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:11697907}.
CC -!- MISCELLANEOUS: Both catalyzed reactions, i.e. dTDP-glucose biosynthesis
CC and pyrophosphorolysis, follow a sequential ordered bi-bi catalytic
CC mechanism. {ECO:0000269|PubMed:11697907}.
CC -!- SIMILARITY: Belongs to the glucose-1-phosphate thymidylyltransferase
CC family. {ECO:0000305}.
CC -!- CAUTION: Most extant K-12 lines do not express O-antigen because of
CC mutations in dTDP-rhamnose biosynthetic genes or in the
CC rhamnosyltransferase gene wbbL. {ECO:0000305}.
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DR EMBL; U09876; AAB88400.1; -; Genomic_DNA.
DR EMBL; U00096; AAC75100.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA15881.1; -; Genomic_DNA.
DR EMBL; U03041; AAC31629.1; -; Genomic_DNA.
DR PIR; F64969; F64969.
DR RefSeq; NP_416543.1; NC_000913.3.
DR RefSeq; WP_000783975.1; NZ_LN832404.1.
DR PDB; 1H5R; X-ray; 1.90 A; A/B/C/D=1-293.
DR PDB; 1H5S; X-ray; 2.30 A; A/B/C/D=1-293.
DR PDB; 1H5T; X-ray; 1.90 A; A/B/C/D=1-293.
DR PDBsum; 1H5R; -.
DR PDBsum; 1H5S; -.
DR PDBsum; 1H5T; -.
DR AlphaFoldDB; P37744; -.
DR SMR; P37744; -.
DR BioGRID; 4259680; 190.
DR IntAct; P37744; 9.
DR STRING; 511145.b2039; -.
DR SWISS-2DPAGE; P37744; -.
DR jPOST; P37744; -.
DR PaxDb; P37744; -.
DR PRIDE; P37744; -.
DR EnsemblBacteria; AAC75100; AAC75100; b2039.
DR EnsemblBacteria; BAA15881; BAA15881; BAA15881.
DR GeneID; 945154; -.
DR KEGG; ecj:JW2024; -.
DR KEGG; eco:b2039; -.
DR PATRIC; fig|1411691.4.peg.212; -.
DR EchoBASE; EB1921; -.
DR eggNOG; COG1209; Bacteria.
DR HOGENOM; CLU_029499_9_0_6; -.
DR InParanoid; P37744; -.
DR OMA; PFIMYLG; -.
DR PhylomeDB; P37744; -.
DR BioCyc; EcoCyc:DTDPGLUCOSEPP-MON; -.
DR BioCyc; MetaCyc:DTDPGLUCOSEPP-MON; -.
DR UniPathway; UPA00124; -.
DR UniPathway; UPA00281; -.
DR EvolutionaryTrace; P37744; -.
DR PRO; PR:P37744; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0008879; F:glucose-1-phosphate thymidylyltransferase activity; IDA:EcoCyc.
DR GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR GO; GO:0046872; F:metal ion binding; IDA:EcoCyc.
DR GO; GO:0019305; P:dTDP-rhamnose biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0045226; P:extracellular polysaccharide biosynthetic process; IEA:InterPro.
DR GO; GO:0009243; P:O antigen biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0051289; P:protein homotetramerization; IDA:EcoCyc.
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; Lipopolysaccharide biosynthesis; Magnesium; Metal-binding;
KW Nucleotidyltransferase; Reference proteome; Transferase.
FT CHAIN 1..293
FT /note="Glucose-1-phosphate thymidylyltransferase 1"
FT /id="PRO_0000207991"
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 CONFLICT 247
FT /note="Q -> P (in Ref. 6; AAC31629)"
FT /evidence="ECO:0000305"
FT STRAND 4..9
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 15..17
FT /evidence="ECO:0007829|PDB:1H5R"
FT TURN 18..22
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 26..28
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 29..31
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 38..46
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 49..56
FT /evidence="ECO:0007829|PDB:1H5R"
FT TURN 58..60
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 61..68
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 72..74
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 77..82
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 91..95
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 97..100
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 105..109
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 113..115
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 119..128
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 131..139
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 143..145
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 146..151
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 153..155
FT /evidence="ECO:0007829|PDB:1H5S"
FT STRAND 157..163
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 170..179
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 183..189
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 194..197
FT /evidence="ECO:0007829|PDB:1H5S"
FT HELIX 200..209
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 213..217
FT /evidence="ECO:0007829|PDB:1H5R"
FT STRAND 222..225
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 230..247
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 254..260
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 266..273
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 274..276
FT /evidence="ECO:0007829|PDB:1H5R"
FT HELIX 280..288
FT /evidence="ECO:0007829|PDB:1H5R"
SQ SEQUENCE 293 AA; 32694 MW; BA895362D1C5CA55 CRC64;
MKMRKGIILA GGSGTRLYPV TMAVSKQLLP IYDKPMIYYP LSTLMLAGIR DILIISTPQD
TPRFQQLLGD GSQWGLNLQY KVQPSPDGLA QAFIIGEEFI GGDDCALVLG DNIFYGHDLP
KLMEAAVNKE SGATVFAYHV NDPERYGVVE FDKNGTAISL EEKPLEPKSN YAVTGLYFYD
NDVVQMAKNL KPSARGELEI TDINRIYLEQ GRLSVAMMGR GYAWLDTGTH QSLIEASNFI
ATIEERQGLK VSCPEEIAFR KGFIDVEQVR KLAVPLIKNN YGQYLYKMTK DSN
