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Literature summary for 2.4.1.212 extracted from

  • Kumari, K.; Tlapak-Simmons, V.L.; Baggenstoss, B.A.; Weigel, P.H.
    The streptococcal hyaluronan synthases are inhibited by sulfhydryl-modifying reagents, but conserved cysteine residues are not essential for enzyme function (2002), J. Biol. Chem., 277, 13943-13951.
    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 as His-tagged proteins in Escherichia coli Streptococcus dysgalactiae subsp. equisimilis

Protein Variants

Protein Variants Comment Organism
C226A site-directed mutagenesis, 24% remaining activity and altered kinetic constants compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C226A/C262A site-directed mutagenesis, 3.2% remaining activity and altered kinetic constants compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C226A/C262A/C367A site-directed mutagenesis, 1.4% remaining activity and altered kinetic constants compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C226A/C281A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C226A/C367A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C226S site-directed mutagenesis, highly reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C262A site-directed mutagenesis, reduced reaction velocity and increased Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C262A/C281A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C262A/C367A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C262S site-directed mutagenesis, reduced reaction velocity and increased Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C281A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C281A/C367A site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C281S site-directed mutagenesis, reduced reaction velocity and altered Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C367A site-directed mutagenesis, increased reaction velocity and Km values compared to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
C367S site-directed mutagenesis, kinetics similar to the wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
additional information construction and kinetic analysis of Cys-deletion mutants, overview Streptococcus dysgalactiae subsp. equisimilis

Inhibitors

Inhibitors Comment Organism Structure
DTNB 52% inhibition at 0.5 mM Streptococcus dysgalactiae subsp. equisimilis
DTNB 52% inhibition at 0.5 mM Streptococcus pyogenes
iodoacetamide 15% inhibition at 5 mM, biphasic inhibition Streptococcus dysgalactiae subsp. equisimilis
iodoacetamide 13% inhibition at 5 mM, biphasic inhibition Streptococcus pyogenes
methylmethanethiosulfonate 93% reduced activity at 0.05 mM Streptococcus dysgalactiae subsp. equisimilis
methylmethanethiosulfonate 89% reduced activity at 0.05 mM Streptococcus pyogenes
N-ethylmaleimide biphasic inhibition, about 50% reduced reaction velocity, unaltered Km-value, 70% inhibition at 5 mM Streptococcus dysgalactiae subsp. equisimilis
N-ethylmaleimide 60% inhibition at 5 mM, biphasic inhibition Streptococcus pyogenes
sodium arsenite 40% inhibition at 10 mM Streptococcus dysgalactiae subsp. equisimilis
sodium arsenite 46% inhibition at 10 mM Streptococcus pyogenes
sulfhydryl reagents enzyme is inhibited by sulfhydryl-modifying reagents, inhibition mode and mechanism Streptococcus dysgalactiae subsp. equisimilis
sulfhydryl reagents enzyme is inhibited by sulfhydryl-modifying reagents, inhibition mode and mechanism Streptococcus pyogenes

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics, kinetic analysis of Cys-deletion mutants, overview Streptococcus dysgalactiae subsp. equisimilis
0.04
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C281A Streptococcus dysgalactiae subsp. equisimilis
0.044
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C226S Streptococcus dysgalactiae subsp. equisimilis
0.053
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A/C281A Streptococcus dysgalactiae subsp. equisimilis
0.056
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C281S Streptococcus dysgalactiae subsp. equisimilis
0.065
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C281A/C367A Streptococcus dysgalactiae subsp. equisimilis
0.074
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
0.077
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant wild-type enzyme Streptococcus dysgalactiae subsp. equisimilis
0.079
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A/C367A Streptococcus dysgalactiae subsp. equisimilis
0.079
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C367S Streptococcus dysgalactiae subsp. equisimilis
0.085
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C367A Streptococcus dysgalactiae subsp. equisimilis
0.088
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C226A Streptococcus dysgalactiae subsp. equisimilis
0.09
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C367A Streptococcus dysgalactiae subsp. equisimilis
0.091
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C367S Streptococcus dysgalactiae subsp. equisimilis
0.096
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C262S Streptococcus dysgalactiae subsp. equisimilis
0.098
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C281S Streptococcus dysgalactiae subsp. equisimilis
0.113
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262A/C281A Streptococcus dysgalactiae subsp. equisimilis
0.121
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262A/C367A Streptococcus dysgalactiae subsp. equisimilis
0.13
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C281A Streptococcus dysgalactiae subsp. equisimilis
0.134
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A/C262A Streptococcus dysgalactiae subsp. equisimilis
0.146
-
 UDP-D-glucuronate pH 7.0, 30°C, recombinant mutant C262A Streptococcus dysgalactiae subsp. equisimilis
0.153
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262S Streptococcus dysgalactiae subsp. equisimilis
0.154
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226A Streptococcus dysgalactiae subsp. equisimilis
0.186
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C262A Streptococcus dysgalactiae subsp. equisimilis
0.232
-
 UDP-N-acetyl-D-glucosamine pH 7.0, 30°C, recombinant mutant C226S Streptococcus dysgalactiae subsp. equisimilis

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane bound Streptococcus dysgalactiae subsp. equisimilis 16020
-
membrane bound Streptococcus pyogenes 16020
-

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 biosynthesis of hyaluronan [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate Streptococcus pyogenes biosynthesis of hyaluronan [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?

Organism

Organism UniProt Comment Textmining
Streptococcus dysgalactiae subsp. equisimilis
-
-
-
Streptococcus pyogenes Q5X9A9
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Streptococcus dysgalactiae subsp. equisimilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information conserved cysteine residues are not essential for enzyme function Streptococcus dysgalactiae subsp. equisimilis ?
-
 ?
additional information conserved cysteine residues are not essential for enzyme function Streptococcus pyogenes ?
-
 ?
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
-
 Streptococcus pyogenes [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate biosynthesis of hyaluronan 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 biosynthesis of hyaluronan Streptococcus pyogenes [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP
-
 ?

Synonyms

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

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 37 assay at Streptococcus dysgalactiae subsp. equisimilis

pH Optimum

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