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Literature summary for 2.4.1.212 extracted from

  • Krupa, J.C.; Shaya, D.; Chi, L.; Linhardt, R.J.; Cygler, M.; Withers, S.G.; Mort, J.S.
    Quantitative continuous assay for hyaluronan synthase (2007), Anal. Biochem., 361, 218-225.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
analysis a rapid, continuous, and convenient three-enzyme coupled UV absorption assay is developed to quantitate the glucuronic acid and N-acetylglucosamine transferase activities of hyaluronan synthase. Activity is measured by coupling the UDP produced from the PmHAS-catalyzed transfer of UDP-GlcNAc and UDP-GlcUA to a hyaluronic acid tetrasaccharide primer with the oxidation of NADH. Using a fluorescently labeled primer, the products are characterized by gel electrophoresis. The assay can be used to determine kinetic parameters, inhibition constants, and mechanistic aspects of this enzyme. In addition, it can be used to quantify PmHAS during purification of the enzyme from culture media Pasteurella multocida

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pasteurella multocida

Protein Variants

Protein Variants Comment Organism
additional information a truncated soluble form of recombinant PmHAS (residues 1–703) can catalyze the glycosyl transfers in a time- and concentration-dependent manner Pasteurella multocida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.014
-
UDP-alpha-D-glucuronate 25°C, pH 7.5 Pasteurella multocida
0.66
-
UDP-alpha-N-acetyl-D-glucosamine 25°C, pH 7.5 Pasteurella multocida
0.91
-
hyaluronic acid tetrasaccharide 25°C, pH 7.5 Pasteurella multocida

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Pasteurella multocida
-
-

Organism

Organism UniProt Comment Textmining
Pasteurella multocida
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pasteurella multocida

Source Tissue

Source Tissue Comment Organism Textmining
culture medium
-
Pasteurella multocida
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
hyaluronic acid tetrasaccharide + UDP-alpha-D-glucuronate
-
Pasteurella multocida ?
-
?
hyaluronic acid tetrasaccharide + UDP-alpha-N-acetyl-D-glucosamine
-
Pasteurella multocida ?
-
?

Synonyms

Synonyms Comment Organism
PmHAS
-
Pasteurella multocida