Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli | Streptococcus dysgalactiae subsp. equisimilis |
Protein Variants | Comment | Organism |
---|---|---|
C226A | site-directed mutagenesis, increased sensitivity to inhibition by NEM | Streptococcus dysgalactiae subsp. equisimilis |
C226A/C262A | site-directed mutagenesis, slightly increased sensitivity to inhibition by NEM, and reduced sensitivity to inhibition by sodium arsenite compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
C226A/C281A | site-directed mutagenesis, reduced sensitivity to inhibition by NEM, and highly reduced sensitivity to inhibition by sodium arsenite compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
C226A/C367A | site-directed mutagenesis, reduced sensitivity to inhibition by NEM, and slightly increased sensitivity to inhibition by sodium arsenite compared to the wild-type | Streptococcus dysgalactiae subsp. equisimilis |
C226S | site-directed mutagenesis, reduced sensitivity to inhibition by NEM | Streptococcus dysgalactiae subsp. equisimilis |
C262A | site-directed mutagenesis, increased sensitivity to inhibition by NEM | Streptococcus dysgalactiae subsp. equisimilis |
C262A/C281A | site-directed mutagenesis, reduced sensitivity to inhibition by NEM, and highly reduced sensitivity to inhibition by sodium arsenite compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
C262A/C367A | site-directed mutagenesis, increased sensitivity to inhibition by NEM, and highly reduced sensitivity to inhibition by sodium arsenite compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
C262S | site-directed mutagenesis, increased sensitivity to inhibition by NEM | Streptococcus dysgalactiae subsp. equisimilis |
C281A | site-directed mutagenesis, highly reduced sensitivity to inhibition by NEM | Streptococcus dysgalactiae subsp. equisimilis |
C281A/C367A | site-directed mutagenesis, reduced sensitivity to inhibition by NEM, and highly reduced sensitivity to inhibition by sodium arsenite compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
C281S | site-directed mutagenesis, increased sensitivity to inhibition by NEM | Streptococcus dysgalactiae subsp. equisimilis |
C367A | site-directed mutagenesis, unaltered inhibition by NEM compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
C367S | site-directed mutagenesis, unaltered inhibition by NEM compared to the wild-type enzyme | Streptococcus dysgalactiae subsp. equisimilis |
additional information | construction of cysteine-deletion mutants deleting either C226, C262, C281, or C367, the mutants are less sensitive or not sensisitive to sodium arsenite inhibition, overview | Streptococcus dysgalactiae subsp. equisimilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
N-ethylmaleimide | reacts with the Cys residues C226, C262, and C281, but not with C367, binding of substrates UDP-N-acetyl-D-glucosamine, UDP-D-glucuronate, or of product UDP protects the enzyme from inhibition by NEM, level of inhibition of wild-type and mutant enzymes, overview | Streptococcus dysgalactiae subsp. equisimilis | |
sodium arsenite | inhibition of wild-type and mutant enzymes, no inhibition of C262 deletion mutant, overview | Streptococcus dysgalactiae subsp. equisimilis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cell membrane | bound, enzyme possesses a cysteine cluster localized at the inner surface of the cell membrane near the substrate/product-binding sites | Streptococcus dysgalactiae subsp. equisimilis | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate | Streptococcus dysgalactiae subsp. equisimilis | - |
[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus dysgalactiae subsp. equisimilis | - |
class I enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography | Streptococcus dysgalactiae subsp. equisimilis |
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] | catalytically relevant Cys226 and Cys262 are located near the UDP-binding site and might be cross-linked, structural and functional role of cysteine residues, overview | Streptococcus dysgalactiae subsp. equisimilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate | - |
Streptococcus dysgalactiae subsp. equisimilis | [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP | - |
? | |
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate | addition of monosaccharides to the linear heteropolysaccharide chain composed of repeating disaccharides | Streptococcus dysgalactiae subsp. equisimilis | [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the membrane-bound enzyme possesses a membrane-localized cysteine cluster near the substrate-binding sites | Streptococcus dysgalactiae subsp. equisimilis |
Synonyms | Comment | Organism |
---|---|---|
HAS | - |
Streptococcus dysgalactiae subsp. equisimilis |
More | the enzyme belongs to the class I hyaluronan synthases | Streptococcus dysgalactiae subsp. equisimilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptococcus dysgalactiae subsp. equisimilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Streptococcus dysgalactiae subsp. equisimilis |