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Literature summary extracted from

  • van Straaten, K.E.; Routier, F.H.; Sanders, D.A.
    Structural insight into the unique substrate binding mechanism and flavin redox state of UDP-galactopyranose mutase from Aspergillus fumigatus (2012), J. Biol. Chem., 287, 10780-10790.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
5.4.99.9 wild-type and mutant enzyme in complex with substrate UDP-alpha-D-galactopyranose or with inhibitor UDP, microbatch method at room temperature, for the substrate complex crystals: 10 mg/ml protein in 50 mM Tris, pH 8.0, 5 mM DTT, with 10 mM UDP-alpha-D-galactopyranose, and 10 mM dithionite, for the inhibitor complex crystals: 10 mg/ml protein in 25 mM Tris malonate, pH 8.0, and 10 mM UDP, for mutant enzyme-substrate complexes: 10 mg/ml protein in 25 mM Tris malonate, pH 8.0, with 15 mM UDP-alpha-D-galactopyranose, mixing of equal volumes of protein solution (with or without ligand) and crystallization solution and overlaid with oil, X-ray diffraction structure determination and analysis at 2.3-3.15 A resolution Aspergillus fumigatus

Protein Variants

EC Number Protein Variants Comment Organism
5.4.99.9 R182A site-directed mutagenesis, the mutant is almost inactive Aspergillus fumigatus
5.4.99.9 R182K site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Aspergillus fumigatus
5.4.99.9 R327A site-directed mutagenesis, inactive mutant Aspergillus fumigatus
5.4.99.9 R327K site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Aspergillus fumigatus

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.4.99.9 UDP
-
 Aspergillus fumigatus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.4.99.9 additional information
-
 additional information steady-state kinetics of wild-type and mutant enzymes, overview Aspergillus fumigatus
5.4.99.9 0.022
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R182K Aspergillus fumigatus
5.4.99.9 0.0425
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, wild-type enzyme Aspergillus fumigatus
5.4.99.9 0.0429
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R327K Aspergillus fumigatus
5.4.99.9 0.607
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R182A Aspergillus fumigatus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.4.99.9 UDP-alpha-D-galactopyranose Aspergillus fumigatus
-
 UDP-alpha-D-galactofuranose
-
 r

Organism

EC Number Organism UniProt Comment Textmining
5.4.99.9 Aspergillus fumigatus Q4W1X2
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.4.99.9 UDP-alpha-D-galactopyranose
-
 Aspergillus fumigatus UDP-alpha-D-galactofuranose
-
 r

Synonyms

EC Number Synonyms Comment Organism
5.4.99.9 UGM
-
 Aspergillus fumigatus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.4.99.9 37
-
 assay at Aspergillus fumigatus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.4.99.9 0.079
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R182A Aspergillus fumigatus
5.4.99.9 0.12
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R327K Aspergillus fumigatus
5.4.99.9 0.44
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R182K Aspergillus fumigatus
5.4.99.9 8.72
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, wild-type enzyme Aspergillus fumigatus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4.99.9 7
-
 assay at Aspergillus fumigatus

Cofactor

EC Number Cofactor Comment Organism Structure
5.4.99.9 flavin the flavin needs to be reduced for the enzyme to be active Aspergillus fumigatus

General Information

EC Number General Information Comment Organism
5.4.99.9 evolution conserved active site residues in Aspegillus fumigatus UGM compared to prokaryotic UGMs, overview Aspergillus fumigatus
5.4.99.9 additional information Arg182 and Arg327 play important roles in stabilizing the position of the diphosphates of the nucleotide sugar and help to facilitate the positioning of the galactose moiety for catalysis. Substrate recognition and structural changes observed upon substrate binding involving the mobile loops and the critical arginine residues Arg182 and Arg327, overview. The Aspergillus fumigatus enzyme contains a third flexible loop (loop III) above the si-face of the isoalloxazine ring that changes position depending on the redox state of the flavin cofactor Aspergillus fumigatus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.4.99.9 0.00013
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R182A Aspergillus fumigatus
5.4.99.9 0.003
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R327K Aspergillus fumigatus
5.4.99.9 0.02
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, mutant R182K Aspergillus fumigatus
5.4.99.9 0.205
-
 UDP-alpha-D-galactopyranose pH 7.0, 37°C, wild-type enzyme Aspergillus fumigatus