Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain BJ5461 | Xenopus laevis |
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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | a nucleotide with 2 phosphate groups and complexed with a Mg2+ ion is absolutely required for activity | Xenopus laevis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Xenopus laevis | - |
isozyme HAS1 | - |
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 |
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 | Comment | Organism |
---|---|---|
HA synthase | - |
Xenopus laevis |
HAS | - |
Xenopus laevis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Xenopus laevis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Xenopus laevis |