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Literature summary extracted from

  • Kotake, T.; Hojo, S.; Tajima, N.; Matsuoka, K.; Koyama, T.; Tsumuraya, Y.
    A bifunctional enzyme with L-fucokinase and GDP-L-fucose pyrophosphorylase activities salvages free L-fucose in Arabidopsis (2008), J. Biol. Chem., 283, 8125-8135.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.52 expression in Escherichia coli with chaperones GroEL, GroES Arabidopsis thaliana
2.7.7.30 expression in BL21 (DE3) gold strain of Escherichia coli Arabidopsis thaliana

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.52 G133A mutation abolishes GDP-L-fucose diphosphorylase activity, hardly affects fucokinase activity Arabidopsis thaliana
2.7.1.52 G830A mutation reduces fucokinase activity by more than 90%, while preserving the GDP-L-fucose diphosphorylase activity Arabidopsis thaliana

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.30 diphosphate addition of diphosphate strongly inhibits the formation of GDP-L-Fuc by rAtFKGPi Arabidopsis thaliana

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.52 0.45
-
 ATP pH 7.5, 35°C Arabidopsis thaliana
2.7.1.52 1
-
 L-fucose pH 7.5, 35°C Arabidopsis thaliana
2.7.7.30 0.052
-
 L-fucose-1-phosphate
-
 Arabidopsis thaliana
2.7.7.30 0.17
-
 GTP
-
 Arabidopsis thaliana

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.52 Mg2+ or Mn2+, absolutely required for fucokinase activity Arabidopsis thaliana
2.7.1.52 Mn2+ or Mg2+, absolutely required for fucokinase activity Arabidopsis thaliana
2.7.7.30 Mg2+
-
 Arabidopsis thaliana
2.7.7.30 Mn2+ has only a poor effect on the GDP-L-Fuc pyrophosphorylase activity of rAtFKGP compared with Mg2+ Arabidopsis thaliana

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.52 116350
-
 x * 116350, calculated, x * 120000, SDS-PAGE Arabidopsis thaliana
2.7.1.52 120000
-
 x * 116350, calculated, x * 120000, SDS-PAGE Arabidopsis thaliana
2.7.7.30 116400
-
 AtFKGP, calculated Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.30 GTP + L-fucose 1-phosphate Arabidopsis thaliana the monosaccharide L-fucose is found as a constituent of cell wall polysaccharides and sugar moieties of glycoproteins and has important physiological functions in seed plants diphosphate + GDP-L-fucose
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.52 Arabidopsis thaliana
-
 bifunctional enzyme, L-fucokinase and GDP-L-fucose diphosphorylase activities
-
2.7.7.30 Arabidopsis thaliana
-
 ecotype Columbia (Col)
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.7.1.52 additional information peptide sequence does not contain a secretory signal or transmembrane domain Arabidopsis thaliana

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.52 recombinant protein with His-tag and fused to thioredoxin, partial purification Arabidopsis thaliana
2.7.7.30 Sepharose FF column, the recombinant protein fused to thioredoxin and His tags is partially purified by chelating chromatography Arabidopsis thaliana

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.52 additional information expression in all tissues examined Arabidopsis thaliana
-
2.7.7.30 cauline leaf mature plant Arabidopsis thaliana
-
2.7.7.30 cotyledon seedling Arabidopsis thaliana
-
2.7.7.30 flower mature plant Arabidopsis thaliana
-
2.7.7.30 flower bud mature plant, relatively high expression of AtFKGP mRNA Arabidopsis thaliana
-
2.7.7.30 inflorescence mature plant Arabidopsis thaliana
-
2.7.7.30 root seedling Arabidopsis thaliana
-
2.7.7.30 rosette leaf mature plant Arabidopsis thaliana
-
2.7.7.30 true leaf seedling Arabidopsis thaliana
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.52 82.9
-
 pH 7.5, 35°C Arabidopsis thaliana
2.7.7.30 0.71
-
-
 Arabidopsis thaliana

Storage Stability

EC Number Storage Stability Organism
2.7.1.52 -20°C, 50% glycerol, 1 mM PMSF, 0.2 M NaCl, 50 mM Tris-HCl buffer, pH 7.5, stable for 1 month Arabidopsis thaliana
2.7.1.52 4°C, 1 day, loss of 50% of GDP-L-fucose diphosphorylase activity Arabidopsis thaliana
2.7.7.30 -20°C, 50% (w/v) glycerol, 1 mM PMSF, 0.2 M NaCl, 50 mM Tris-HCl buffer, pH 7.5, 1 month, no loss of activity Arabidopsis thaliana
2.7.7.30 4°C, 1mM PMSF, 0.2 M NaCl, 50 mM Tris-HCl buffer, pH 7.5, 1 day, loss of more than 50% activity Arabidopsis thaliana

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.52 ATP + L-fucose
-
 Arabidopsis thaliana ADP + beta-L-fucose 1-phosphate
-
 ?
2.7.1.52 additional information enzyme generates GDP-L-fucose from L-fucose 1-phosphate and GTP. No substrates for fucokinase activity are: L-arabinose, glucose, galactose, mannose, xylose, L-rhamnose, galactosamine, glucosamine, N-acetylglucosamine Arabidopsis thaliana ?
-
 ?
2.7.7.30 GTP + L-fucose 1-phosphate the monosaccharide L-fucose is found as a constituent of cell wall polysaccharides and sugar moieties of glycoproteins and has important physiological functions in seed plants Arabidopsis thaliana diphosphate + GDP-L-fucose
-
 ?
2.7.7.30 GTP + L-fucose 1-phosphate strict substrate specificity, no activity with L-Ara-1-P, Gal-1-P, Glc-1-P, Man-1-P, Xyl-1-P, GlcA-1-P, and Glc-NAc-1-P as glycosyl donor, and it uses specifically GTP as the nucleotide donor Arabidopsis thaliana diphosphate + GDP-L-fucose
-
 ?
2.7.7.30 L-fucose-1-phosphate + GTP
-
 Arabidopsis thaliana GDP-L-fucose + diphosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.52 ? x * 116350, calculated, x * 120000, SDS-PAGE Arabidopsis thaliana

Synonyms

EC Number Synonyms Comment Organism
2.7.1.52 Fkgp
-
 Arabidopsis thaliana
2.7.7.30 AtFKGP bifunctional enzyme with L-fucokinase and GDP-L-Fuc pyrophosphorylase Arabidopsis thaliana
2.7.7.30 GDP-L-Fuc pyrophosphorylase
-
 Arabidopsis thaliana
2.7.7.30 GDP-L-fucose pyrophosphorylase
-
 Arabidopsis thaliana

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.52 40
-
 optimum for fucokinase activity Arabidopsis thaliana

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.30 30 45 rAtFKGP, GDP-L-Fuc pyrophosphorylase activity Arabidopsis thaliana

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.1.52 50
-
 loss of 99% of activity Arabidopsis thaliana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.52 0.34
-
 L-fucose pH 7.5, 35°C Arabidopsis thaliana
2.7.1.52 0.38
-
 ATP pH 7.5, 35°C Arabidopsis thaliana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.52 10.5
-
 optimum for fucokinase activity Arabidopsis thaliana

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.30 6.5 8
-
 Arabidopsis thaliana

pI Value

EC Number Organism Comment pI Value Maximum pI Value
2.7.1.52 Arabidopsis thaliana calculated
-
 5.9
2.7.7.30 Arabidopsis thaliana calculated, theoretical isoelectric point
-
 5.87