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Literature summary for 2.4.1.212 extracted from

  • Pummill, P.E.; DeAngelis, P.L.
    Evaluation of critical structural elements of UDP-sugar substrates and certain cysteine residues of a vertebrate hyaluronan synthase (2002), J. Biol. Chem., 277, 21610-21616.
    View publication on PubMed
Search for more literature for 2.4.1.212

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain BJ5461 Xenopus laevis

Protein Variants

Protein Variants Comment Organism
C117F site-directed mutagenesis, no expression in yeast possible Xenopus laevis
C117L site-directed mutagenesis, reduced activity compared to the wild-type enzyme Xenopus laevis
C117S site-directed mutagenesis, activity is similar to the wild-type enzyme Xenopus laevis
C210S site-directed mutagenesis, reduced activity compared to the wild-type enzyme Xenopus laevis
C239S site-directed mutagenesis, activity is similar to the wild-type enzyme Xenopus laevis
C239S/C337S site-directed mutagenesis, reduced recombinant expression level, activity is similar to the wild-type enzyme Xenopus laevis
C298F site-directed mutagenesis, poor recombinant expression level, highly reduced activity compared to the wild-type enzyme Xenopus laevis
C298L site-directed mutagenesis, poor recombinant expression level, highly reduced activity compared to the wild-type enzyme Xenopus laevis
C298S site-directed mutagenesis, activity is similar to the wild-type enzyme Xenopus laevis
C304S site-directed mutagenesis, activity is similar to the wild-type enzyme Xenopus laevis
C304S/C337S site-directed mutagenesis, reduced activity compared to the wild-type enzyme Xenopus laevis
C307S site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Xenopus laevis
C307S/C337S site-directed mutagenesis, reduced recombinant expression level, inactive mutant Xenopus laevis
C337S site-directed mutagenesis, increased Km for UDP-N-acetylglucosamine compared to the wild-type enzyme Xenopus laevis

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
 Xenopus laevis
N-ethylmaleimide inhibits wild-type and mutant enzymes by about 90% at 0.2 mM, UDP-N-acetyl-D-glucosamine protects the enzyme best against inactivation by NEM compared to diverse other nucleotide compounds, overview Xenopus laevis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Km values of the enzyme in standard reaction in presence of diverse protecting nucleotide compounds, overview Xenopus laevis
0.11
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant wild-type enzyme at low UDP-N-acetyl-D-glucosamine concentration, and mutant C239S Xenopus laevis
0.12
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant mutants C117S and C210S Xenopus laevis
0.16
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant mutant C298S Xenopus laevis
0.18
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant mutant C304S Xenopus laevis
0.19
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant wild-type enzyme Xenopus laevis
0.26
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant wild-type enzyme at low UDP-glucuronate concentration Xenopus laevis
0.32
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C239S Xenopus laevis
0.34
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C117S Xenopus laevis
0.4
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant wild-type enzyme and mutants C210S, C337S at low UDP-glucuronate concentration Xenopus laevis
0.43
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C298S Xenopus laevis
0.47
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C304S Xenopus laevis
0.7
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant mutant C337S Xenopus laevis
0.88
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C337S at higher UDP-glucuronate concentration Xenopus laevis
0.89
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant mutant C304S/C337S Xenopus laevis
0.93
-
 UDP-D-glucuronate pH 7.5, 30°C, recombinant mutant C239S/C337S Xenopus laevis
1
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C239S/C337S Xenopus laevis
1.1
-
 UDP-N-acetyl-D-glucosamine pH 7.5, 30°C, recombinant mutant C304S/C337S Xenopus laevis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ a nucleotide with 2 phosphate groups and complexed with a Mg2+ ion is absolutely required for activity Xenopus laevis

Organism

Organism UniProt Comment Textmining
Xenopus laevis
-
 isozyme HAS1
-

Reaction

Reaction Comment Organism Reaction ID
UDP-N-acetyl-alpha-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] = UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] Cys307 might be a catalytic residue or be involved in maintaining structure, Cys337 is involved in UDP-sugar substrate binding, enzyme amino acid residues involved in interactions with the substrate, overview Xenopus laevis
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the substrate binding selectivity is more relaxed than the specificity of catalytic transfer, a nucleotide with 2 phosphate groups and complexed with a Mg2+ ion is absolutely required for activity Xenopus laevis ?
-
 ?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate addition of monosaccharides to the linear heteropolysaccharide chain Xenopus laevis [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?

Synonyms

Synonyms Comment Organism
HA synthase
-
 Xenopus laevis
HAS
-
 Xenopus laevis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Xenopus laevis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Xenopus laevis