Skip Header

You are using a version of Internet Explorer that may not display all features of this website. Please upgrade to a modern browser.
Contribute Send feedback
Read comments (?) or add your own

P32055 (FCL_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 124. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (4) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
GDP-L-fucose synthase

EC=1.1.1.271
Alternative name(s):
GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase
Gene names
Name:fcl
Synonyms:wcaG, yefB
Ordered Locus Names:b2052, JW2037
OrganismEscherichia coli (strain K12) [Reference proteome] [HAMAP]
Taxonomic identifier83333 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length321 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Catalyzes the two-step NADP-dependent conversion of GDP-4-dehydro-6-deoxy-D-mannose to GDP-fucose, involving an epimerase and a reductase reaction. Ref.6 Ref.7 Ref.10

Catalytic activity

GDP-beta-L-fucose + NADP+ = GDP-4-dehydro-6-deoxy-alpha-D-mannose + NADPH. Ref.6 Ref.7 Ref.10

Enzyme regulation

Subject to product inhibition by NADP and GDP-fucose. Ref.7

Pathway

Nucleotide-sugar biosynthesis; GDP-L-fucose biosynthesis via de novo pathway; GDP-L-fucose from GDP-alpha-D-mannose: step 2/2. Ref.6

Exopolysaccharide biosynthesis; colanic acid biosynthesis. Ref.6

Subunit structure

Homodimer. Ref.8

Subcellular location

Cytoplasm HAMAP-Rule MF_00956.

Sequence similarities

Belongs to the NAD(P)-dependent epimerase/dehydratase family. Fucose synthase subfamily.

Biophysicochemical properties

Kinetic parameters:

KM=9 µM for NADPH Ref.7 Ref.10

KM=109 µM for NADH

KM=29 µM for GDP-4-keto-6-deoxymannose

Vmax=363.5 µmol/min/mg enzyme

pH dependence:

Optimum pH is 6-6.5.

Ontologies

Keywords
   Cellular componentCytoplasm
   LigandNADP
   Molecular functionIsomerase
Oxidoreductase
   Technical term3D-structure
Complete proteome
Multifunctional enzyme
Reference proteome
Gene Ontology (GO)
   Biological_process'de novo' GDP-L-fucose biosynthetic process

Inferred from electronic annotation. Source: UniProtKB-HAMAP

colanic acid biosynthetic process

Inferred from electronic annotation. Source: UniProtKB-UniPathway

   Cellular_componentcytoplasm

Inferred from electronic annotation. Source: UniProtKB-SubCell

   Molecular_functionGDP-L-fucose synthase activity

Inferred from direct assay Ref.7. Source: EcoCyc

NADP+ binding

Inferred from electronic annotation. Source: UniProtKB-HAMAP

isomerase activity

Inferred from electronic annotation. Source: UniProtKB-KW

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 321321GDP-L-fucose synthase HAMAP-Rule MF_00956
PRO_0000174358

Regions

Nucleotide binding10 – 167NADP HAMAP-Rule MF_00956
Nucleotide binding36 – 416NADP HAMAP-Rule MF_00956
Nucleotide binding105 – 1084NADP HAMAP-Rule MF_00956
Nucleotide binding163 – 1664NADP HAMAP-Rule MF_00956

Sites

Active site1361Proton donor/acceptor Ref.10
Binding site1401NADP
Binding site1791NADP
Binding site1871Substrate Probable
Binding site2021Substrate By similarity
Binding site2091Substrate By similarity
Binding site2781Substrate By similarity
Site1071Important for catalytic activity
Site1091Important for catalytic activity
Site1401Lowers pKa of active site Tyr

Experimental info

Mutagenesis1071S → A: Nearly abolishes catalytic activity. Minor effect of affinity for NADPH and substrate. Ref.10
Mutagenesis1091C → A: Nearly abolishes catalytic activity.
Mutagenesis1361Y → E: Abolishes enzyme activity. Ref.10
Mutagenesis1401K → R: Reduces catalytic activity 20-fold. Ref.10
Mutagenesis1401K → S: Nearly abolishes catalytic activity. Ref.10
Mutagenesis1791H → N: Nearly abolishes catalytic activity. Ref.10
Mutagenesis1871R → A: Decreases affinity for the substrate GDP-4-keto-6-deoxymannose. Ref.10
Sequence conflict255 – 2562EL → DV no nucleotide entry Ref.1
Sequence conflict255 – 2562EL → DV in AAC77843. Ref.2

Secondary structure

...................................................... 321
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P32055 [UniParc].

Last modified November 1, 1997. Version 2.
Checksum: 97077193D79684C7

FASTA32136,141
        10         20         30         40         50         60 
MSKQRVFIAG HRGMVGSAIR RQLEQRGDVE LVLRTRDELN LLDSRAVHDF FASERIDQVY 

        70         80         90        100        110        120 
LAAAKVGGIV ANNTYPADFI YQNMMIESNI IHAAHQNDVN KLLFLGSSCI YPKLAKQPMA 

       130        140        150        160        170        180 
ESELLQGTLE PTNEPYAIAK IAGIKLCESY NRQYGRDYRS VMPTNLYGPH DNFHPSNSHV 

       190        200        210        220        230        240 
IPALLRRFHE ATAQNAPDVV VWGSGTPMRE FLHVDDMAAA SIHVMELAHE VWLENTQPML 

       250        260        270        280        290        300 
SHINVGTGVD CTIRELAQTI AKVVGYKGRV VFDASKPDGT PRKLLDVTRL HQLGWYHEIS 

       310        320 
LEAGLASTYQ WFLENQDRFR G 

« Hide

References

« Hide 'large scale' references
[1]"Evidence for effect of random genetic drift on G+C content after lateral transfer of fucose pathway genes to Escherichia coli K-12."
Aoyama K., Haase A.M., Reeves P.R.
Mol. Biol. Evol. 11:829-838(1994) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
Strain: K12.
[2]"Organization of the Escherichia coli K-12 gene cluster responsible for production of the extracellular polysaccharide colanic acid."
Stevenson G., Andrianopoulos K., Hobbs M., Reeves P.R.
J. Bacteriol. 178:4885-4893(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
Strain: K12.
[3]"A 460-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 40.1-50.0 min region on the linkage map."
Itoh T., Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., Kasai H., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T., Mizobuchi K., Mori H., Mori T., Motomura K., Nakade S. expand/collapse author list , Nakamura Y., Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y., Sivasundaram S., Tagami H., Takeda J., Takemoto K., Wada C., Yamamoto Y., Horiuchi T.
DNA Res. 3:379-392(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[4]"The complete genome sequence of Escherichia coli K-12."
Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.
Science 277:1453-1462(1997) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / MG1655 / ATCC 47076.
[5]"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."
Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.
Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[6]"Identification of the fucose synthetase gene in the colanic acid gene cluster of Escherichia coli K-12."
Andrianopoulos K., Wang L., Reeves P.R.
J. Bacteriol. 180:998-1001(1998) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, CATALYTIC ACTIVITY, PATHWAY.
[7]"Stereochemical course and steady state mechanism of the reaction catalyzed by the GDP-fucose synthetase from Escherichia coli."
Menon S., Stahl M., Kumar R., Xu G.Y., Sullivan F.
J. Biol. Chem. 274:26743-26750(1999) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES.
[8]"GDP-4-keto-6-deoxy-D-mannose epimerase/reductase from Escherichia coli, a key enzyme in the biosynthesis of GDP-L-fucose, displays the structural characteristics of the RED protein homology superfamily."
Rizzi M., Tonetti M., Vigevani P., Sturla L., Bisso A., Flora A.D., Bordo D., Bolognesi M.
Structure 6:1453-1465(1998) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH NADP, SUBUNIT.
[9]"GDP-fucose synthetase from Escherichia coli: structure of a unique member of the short-chain dehydrogenase/reductase family that catalyzes two distinct reactions at the same active site."
Somers W.S., Stahl M.L., Sullivan F.X.
Structure 6:1601-1612(1998) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH NADP.
Strain: K12.
[10]"Probing the catalytic mechanism of GDP-4-keto-6-deoxy-d-mannose Epimerase/Reductase by kinetic and crystallographic characterization of site-specific mutants."
Rosano C., Bisso A., Izzo G., Tonetti M., Sturla L., De Flora A., Bolognesi M.
J. Mol. Biol. 303:77-91(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.45 ANGSTROMS) IN COMPLEX WITH NADP, CATALYTIC ACTIVITY, FUNCTION, PREDICTED SUBSTRATE-BINDING SITES, MUTAGENESIS OF SER-107; TYR-136; LYS-140; HIS-179 AND ARG-187, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITE.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
U38473 Genomic DNA. Translation: AAC77843.1.
U00096 Genomic DNA. Translation: AAC75113.1.
AP009048 Genomic DNA. Translation: BAA15908.1.
PIRC64971.
RefSeqNP_416556.1. NC_000913.3.
YP_490294.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1BSVX-ray2.20A1-321[»]
1BWSX-ray2.20A1-321[»]
1E6UX-ray1.45A3-321[»]
1E7QX-ray1.60A3-321[»]
1E7RX-ray1.60A1-321[»]
1E7SX-ray1.50A1-321[»]
1FXSX-ray2.30A1-321[»]
1GFSX-ray2.20A1-321[»]
ProteinModelPortalP32055.
SMRP32055. Positions 3-319.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

IntActP32055. 2 interactions.
STRING511145.b2052.

Proteomic databases

PaxDbP32055.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAC75113; AAC75113; b2052.
BAA15908; BAA15908; BAA15908.
GeneID12930697.
946563.
KEGGecj:Y75_p2015.
eco:b2052.
PATRIC32119437. VBIEscCol129921_2129.

Organism-specific databases

EchoBASEEB1736.
EcoGeneEG11788. fcl.

Phylogenomic databases

eggNOGCOG0451.
HOGENOMHOG000168011.
KOK02377.
OMAIHCAGRV.
OrthoDBEOG6JB132.
PhylomeDBP32055.
ProtClustDBCLSK880275.

Enzyme and pathway databases

BioCycEcoCyc:FCL-MONOMER.
ECOL316407:JW2037-MONOMER.
MetaCyc:FCL-MONOMER.
UniPathwayUPA00128; UER00191.
UPA00980.

Gene expression databases

GenevestigatorP32055.

Family and domain databases

Gene3D3.40.50.720. 2 hits.
HAMAPMF_00956. GDP_fucose_synth.
InterProIPR001509. Epimerase_deHydtase.
IPR028614. GDP_fucose_synth.
IPR016040. NAD(P)-bd_dom.
[Graphical view]
PfamPF01370. Epimerase. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP32055.
PROP32055.

Entry information

Entry nameFCL_ECOLI
AccessionPrimary (citable) accession number: P32055
Secondary accession number(s): P76382
Entry history
Integrated into UniProtKB/Swiss-Prot: October 1, 1993
Last sequence update: November 1, 1997
Last modified: April 16, 2014
This is version 124 of the entry and version 2 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways

Escherichia coli

Escherichia coli (strain K12): entries and cross-references to EcoGene