FCL_ECOLI
ID FCL_ECOLI Reviewed; 321 AA.
AC P32055; P76382;
DT 01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1997, sequence version 2.
DT 03-AUG-2022, entry version 173.
DE RecName: Full=GDP-L-fucose synthase {ECO:0000255|HAMAP-Rule:MF_00956};
DE EC=1.1.1.271 {ECO:0000255|HAMAP-Rule:MF_00956, ECO:0000269|PubMed:10480878, ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9473059};
DE AltName: Full=GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase {ECO:0000255|HAMAP-Rule:MF_00956};
GN Name=fcl {ECO:0000255|HAMAP-Rule:MF_00956}; Synonyms=wcaG, yefB;
GN OrderedLocusNames=b2052, JW2037;
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;
RX PubMed=7815923; DOI=10.1093/oxfordjournals.molbev.a040166;
RA Aoyama K., Haase A.M., Reeves P.R.;
RT "Evidence for effect of random genetic drift on G+C content after lateral
RT transfer of fucose pathway genes to Escherichia coli K-12.";
RL Mol. Biol. Evol. 11:829-838(1994).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RX PubMed=8759852; DOI=10.1128/jb.178.16.4885-4893.1996;
RA Stevenson G., Andrianopoulos K., Hobbs M., Reeves P.R.;
RT "Organization of the Escherichia coli K-12 gene cluster responsible for
RT production of the extracellular polysaccharide colanic acid.";
RL J. Bacteriol. 178:4885-4893(1996).
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 FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=9473059; DOI=10.1128/jb.180.4.998-1001.1998;
RA Andrianopoulos K., Wang L., Reeves P.R.;
RT "Identification of the fucose synthetase gene in the colanic acid gene
RT cluster of Escherichia coli K-12.";
RL J. Bacteriol. 180:998-1001(1998).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=10480878; DOI=10.1074/jbc.274.38.26743;
RA Menon S., Stahl M., Kumar R., Xu G.Y., Sullivan F.;
RT "Stereochemical course and steady state mechanism of the reaction catalyzed
RT by the GDP-fucose synthetase from Escherichia coli.";
RL J. Biol. Chem. 274:26743-26750(1999).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH NADP, AND SUBUNIT.
RX PubMed=9817848; DOI=10.1016/s0969-2126(98)00144-0;
RA Rizzi M., Tonetti M., Vigevani P., Sturla L., Bisso A., Flora A.D.,
RA Bordo D., Bolognesi M.;
RT "GDP-4-keto-6-deoxy-D-mannose epimerase/reductase from Escherichia coli, a
RT key enzyme in the biosynthesis of GDP-L-fucose, displays the structural
RT characteristics of the RED protein homology superfamily.";
RL Structure 6:1453-1465(1998).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH NADP.
RC STRAIN=K12;
RX PubMed=9862812; DOI=10.1016/s0969-2126(98)00157-9;
RA Somers W.S., Stahl M.L., Sullivan F.X.;
RT "GDP-fucose synthetase from Escherichia coli: structure of a unique member
RT of the short-chain dehydrogenase/reductase family that catalyzes two
RT distinct reactions at the same active site.";
RL Structure 6:1601-1612(1998).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (1.45 ANGSTROMS) IN COMPLEX WITH NADP, CATALYTIC
RP ACTIVITY, FUNCTION, PREDICTED SUBSTRATE-BINDING SITES, MUTAGENESIS OF
RP SER-107; TYR-136; LYS-140; HIS-179 AND ARG-187, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND ACTIVE SITE.
RX PubMed=11021971; DOI=10.1006/jmbi.2000.4106;
RA Rosano C., Bisso A., Izzo G., Tonetti M., Sturla L., De Flora A.,
RA Bolognesi M.;
RT "Probing the catalytic mechanism of GDP-4-keto-6-deoxy-d-mannose
RT Epimerase/Reductase by kinetic and crystallographic characterization of
RT site-specific mutants.";
RL J. Mol. Biol. 303:77-91(2000).
CC -!- FUNCTION: Catalyzes the two-step NADP-dependent conversion of GDP-4-
CC dehydro-6-deoxy-D-mannose to GDP-fucose, involving an epimerase and a
CC reductase reaction. {ECO:0000255|HAMAP-Rule:MF_00956,
CC ECO:0000269|PubMed:10480878, ECO:0000269|PubMed:11021971,
CC ECO:0000269|PubMed:9473059}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GDP-beta-L-fucose + NADP(+) = GDP-4-dehydro-alpha-D-rhamnose +
CC H(+) + NADPH; Xref=Rhea:RHEA:18885, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57273, ChEBI:CHEBI:57783, ChEBI:CHEBI:57964,
CC ChEBI:CHEBI:58349; EC=1.1.1.271; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_00956, ECO:0000269|PubMed:10480878,
CC ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9473059};
CC -!- ACTIVITY REGULATION: Subject to product inhibition by NADP and GDP-
CC fucose. {ECO:0000269|PubMed:10480878}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=9 uM for NADPH {ECO:0000269|PubMed:10480878,
CC ECO:0000269|PubMed:11021971};
CC KM=109 uM for NADH {ECO:0000269|PubMed:10480878,
CC ECO:0000269|PubMed:11021971};
CC KM=29 uM for GDP-4-keto-6-deoxymannose {ECO:0000269|PubMed:10480878,
CC ECO:0000269|PubMed:11021971};
CC Vmax=363.5 umol/min/mg enzyme {ECO:0000269|PubMed:10480878,
CC ECO:0000269|PubMed:11021971};
CC pH dependence:
CC Optimum pH is 6-6.5. {ECO:0000269|PubMed:10480878,
CC ECO:0000269|PubMed:11021971};
CC -!- PATHWAY: Nucleotide-sugar biosynthesis; GDP-L-fucose biosynthesis via
CC de novo pathway; GDP-L-fucose from GDP-alpha-D-mannose: step 2/2.
CC {ECO:0000255|HAMAP-Rule:MF_00956, ECO:0000269|PubMed:9473059}.
CC -!- PATHWAY: Exopolysaccharide biosynthesis; colanic acid biosynthesis.
CC {ECO:0000269|PubMed:9473059}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11021971,
CC ECO:0000269|PubMed:9817848, ECO:0000269|PubMed:9862812}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- SIMILARITY: Belongs to the NAD(P)-dependent epimerase/dehydratase
CC family. Fucose synthase subfamily. {ECO:0000255|HAMAP-Rule:MF_00956}.
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DR EMBL; U38473; AAC77843.1; -; Genomic_DNA.
DR EMBL; U00096; AAC75113.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA15908.1; -; Genomic_DNA.
DR PIR; C64971; C64971.
DR RefSeq; NP_416556.1; NC_000913.3.
DR RefSeq; WP_000043654.1; NZ_LN832404.1.
DR PDB; 1BSV; X-ray; 2.20 A; A=1-321.
DR PDB; 1BWS; X-ray; 2.20 A; A=1-321.
DR PDB; 1E6U; X-ray; 1.45 A; A=1-321.
DR PDB; 1E7Q; X-ray; 1.60 A; A=1-321.
DR PDB; 1E7R; X-ray; 1.60 A; A=1-321.
DR PDB; 1E7S; X-ray; 1.50 A; A=1-321.
DR PDB; 1FXS; X-ray; 2.30 A; A=1-321.
DR PDB; 1GFS; X-ray; 2.20 A; A=1-321.
DR PDBsum; 1BSV; -.
DR PDBsum; 1BWS; -.
DR PDBsum; 1E6U; -.
DR PDBsum; 1E7Q; -.
DR PDBsum; 1E7R; -.
DR PDBsum; 1E7S; -.
DR PDBsum; 1FXS; -.
DR PDBsum; 1GFS; -.
DR AlphaFoldDB; P32055; -.
DR SMR; P32055; -.
DR BioGRID; 4259691; 221.
DR IntAct; P32055; 2.
DR STRING; 511145.b2052; -.
DR DrugBank; DB02897; Acetylphosphate.
DR DrugBank; DB03461; Nicotinamide adenine dinucleotide phosphate.
DR PaxDb; P32055; -.
DR PRIDE; P32055; -.
DR EnsemblBacteria; AAC75113; AAC75113; b2052.
DR EnsemblBacteria; BAA15908; BAA15908; BAA15908.
DR GeneID; 946563; -.
DR KEGG; ecj:JW2037; -.
DR KEGG; eco:b2052; -.
DR PATRIC; fig|1411691.4.peg.199; -.
DR EchoBASE; EB1736; -.
DR eggNOG; COG0451; Bacteria.
DR HOGENOM; CLU_007383_18_0_6; -.
DR InParanoid; P32055; -.
DR OMA; IHCAGRV; -.
DR PhylomeDB; P32055; -.
DR BioCyc; EcoCyc:FCL-MON; -.
DR BioCyc; MetaCyc:FCL-MON; -.
DR BRENDA; 1.1.1.271; 2026.
DR SABIO-RK; P32055; -.
DR UniPathway; UPA00128; UER00191.
DR UniPathway; UPA00980; -.
DR EvolutionaryTrace; P32055; -.
DR PRO; PR:P32055; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0050577; F:GDP-L-fucose synthase activity; IDA:EcoCyc.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0070401; F:NADP+ binding; IEA:UniProtKB-UniRule.
DR GO; GO:0042351; P:'de novo' GDP-L-fucose biosynthetic process; IEA:UniProtKB-UniRule.
DR GO; GO:0009242; P:colanic acid biosynthetic process; IEA:UniProtKB-UniPathway.
DR CDD; cd05239; GDP_FS_SDR_e; 1.
DR HAMAP; MF_00956; GDP_fucose_synth; 1.
DR InterPro; IPR001509; Epimerase_deHydtase.
DR InterPro; IPR028614; GDP_fucose/colitose_synth.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF01370; Epimerase; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Isomerase; Multifunctional enzyme; NADP;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..321
FT /note="GDP-L-fucose synthase"
FT /id="PRO_0000174358"
FT ACT_SITE 136
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971"
FT BINDING 10..16
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT ECO:0000269|PubMed:9862812"
FT BINDING 36..41
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT ECO:0000269|PubMed:9862812"
FT BINDING 105..108
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT ECO:0000269|PubMed:9862812"
FT BINDING 140
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT ECO:0000269|PubMed:9862812"
FT BINDING 163..166
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT ECO:0000269|PubMed:9862812"
FT BINDING 179
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT ECO:0000269|PubMed:9862812"
FT BINDING 187
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 202
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956"
FT BINDING 209
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956"
FT BINDING 278
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00956"
FT SITE 107
FT /note="Important for catalytic activity"
FT SITE 109
FT /note="Important for catalytic activity"
FT SITE 140
FT /note="Lowers pKa of active site Tyr"
FT MUTAGEN 107
FT /note="S->A: Nearly abolishes catalytic activity. Minor
FT effect of affinity for NADPH and substrate."
FT /evidence="ECO:0000269|PubMed:11021971"
FT MUTAGEN 109
FT /note="C->A: Nearly abolishes catalytic activity."
FT MUTAGEN 136
FT /note="Y->E: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:11021971"
FT MUTAGEN 140
FT /note="K->R: Reduces catalytic activity 20-fold."
FT /evidence="ECO:0000269|PubMed:11021971"
FT MUTAGEN 140
FT /note="K->S: Nearly abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:11021971"
FT MUTAGEN 179
FT /note="H->N: Nearly abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:11021971"
FT MUTAGEN 187
FT /note="R->A: Decreases affinity for the substrate GDP-4-
FT keto-6-deoxymannose."
FT /evidence="ECO:0000269|PubMed:11021971"
FT CONFLICT 255..256
FT /note="EL -> DV (in Ref. 1; no nucleotide entry and 2;
FT AAC77843)"
FT /evidence="ECO:0000305"
FT STRAND 4..9
FT /evidence="ECO:0007829|PDB:1E6U"
FT TURN 10..12
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 14..23
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 29..32
FT /evidence="ECO:0007829|PDB:1E6U"
FT TURN 36..38
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 44..54
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 57..61
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 69..74
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 76..96
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 101..105
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 108..110
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 117..119
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 121..123
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 131..133
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 134..154
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 157..163
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 165..168
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 180..194
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 197..203
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 211..213
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 214..226
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 229..234
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 243..246
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 253..264
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 268..273
FT /evidence="ECO:0007829|PDB:1E6U"
FT STRAND 281..283
FT /evidence="ECO:0007829|PDB:1BSV"
FT HELIX 288..292
FT /evidence="ECO:0007829|PDB:1E6U"
FT HELIX 301..314
FT /evidence="ECO:0007829|PDB:1E6U"
SQ SEQUENCE 321 AA; 36141 MW; 97077193D79684C7 CRC64;
MSKQRVFIAG HRGMVGSAIR RQLEQRGDVE LVLRTRDELN LLDSRAVHDF FASERIDQVY
LAAAKVGGIV ANNTYPADFI YQNMMIESNI IHAAHQNDVN KLLFLGSSCI YPKLAKQPMA
ESELLQGTLE PTNEPYAIAK IAGIKLCESY NRQYGRDYRS VMPTNLYGPH DNFHPSNSHV
IPALLRRFHE ATAQNAPDVV VWGSGTPMRE FLHVDDMAAA SIHVMELAHE VWLENTQPML
SHINVGTGVD CTIRELAQTI AKVVGYKGRV VFDASKPDGT PRKLLDVTRL HQLGWYHEIS
LEAGLASTYQ WFLENQDRFR G
