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Literature summary for 2.4.1.212 extracted from

  • Kumari, K.; Weigel, P.H.
    Identification of a membrane-localized cysteine cluster near the substrate-binding sites of the Streptococcus equisimilis hyaluronan synthase (2005), Glycobiology, 15, 529-539.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged wild-type and mutant enzymes in Escherichia coli Streptococcus dysgalactiae subsp. equisimilis

Protein Variants

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

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

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
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-

Natural Substrates/ Products (Substrates)

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
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[beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
?

Organism

Organism UniProt Comment Textmining
Streptococcus dysgalactiae subsp. equisimilis
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class I enzyme
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Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography Streptococcus dysgalactiae subsp. equisimilis

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] 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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate
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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

Subunits Comment Organism
More the membrane-bound enzyme possesses a membrane-localized cysteine cluster near the substrate-binding sites Streptococcus dysgalactiae subsp. equisimilis

Synonyms

Synonyms Comment Organism
HAS
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Streptococcus dysgalactiae subsp. equisimilis
More the enzyme belongs to the class I hyaluronan synthases Streptococcus dysgalactiae subsp. equisimilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Streptococcus dysgalactiae subsp. equisimilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Streptococcus dysgalactiae subsp. equisimilis