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Literature summary for 2.4.1.212 extracted from

  • Jing, W.; DeAngelis, P.L.
    Analysis of the two active sites of the hyaluronan synthase and the chondroitin synthase of Pasteurella multocida (2003), Glycobiology, 13, 661-671.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutants enzymes in Escherichia coli Pasteurella multocida

Protein Variants

Protein Variants Comment Organism
D247E site-directed mutagenesis, mutant possesses only GlcUA-transferase activity Pasteurella multocida
D247K site-directed mutagenesis, mutant possesses only GlcUA-transferase activity Pasteurella multocida
D247N site-directed mutagenesis, mutant possesses only GlcUA-transferase activity Pasteurella multocida
D249E site-directed mutagenesis, mutant possesses only GlcUA-transferase activity Pasteurella multocida
D249K site-directed mutagenesis, mutant possesses only GlcUA-transferase activity Pasteurella multocida
D249N site-directed mutagenesis, mutant possesses only GlcUA-transferase activity Pasteurella multocida
D370E site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low hyaluronan synthase activity Pasteurella multocida
D370K site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity Pasteurella multocida
D370N site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity Pasteurella multocida
D527E site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity Pasteurella multocida
D527K site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity Pasteurella multocida
D527N site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity Pasteurella multocida
D529E site-directed mutagenesis, mutant possesses GlcNAc-transferase activity, and low hyaluronan synthase activity Pasteurella multocida
D529K site-directed mutagenesis, mutant possesses GlcNAc-transferase activity, and very low hyaluronan synthase activity Pasteurella multocida
D529N site-directed mutagenesis, mutant possesses only GlcNAc-transferase activity Pasteurella multocida
E369D site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity Pasteurella multocida
E369H site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity Pasteurella multocida
E369Q site-directed mutagenesis, mutant possesses GlcUA-transferase activity, and very low GlcNAc-transferase activity Pasteurella multocida
additional information activity rescue of mutants with high substrate concentrations, overview, deletion of residues 1-117 does not affect polymerization activity, construction of different chimeric mutant enzymes comprising residues from Pasteurella multocida type A enzyme and residues of a Pasteurella multocida type F chondroitin synthase, producing an unsulfated chondroitin capsule, the chimeric mutants show different percentages of hyaluronan and chondroitin synthase ativities, overview Pasteurella multocida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics, wild-type and mutant enzymes Pasteurella multocida

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 18% and 39% of the wild-type GlcUA-transferase and GlcNAc-transferase activity with Mn2+, respectively, 0.2 mM Pasteurella multocida
Mg2+ 66% and 77% of the wild-type GlcUA-transferase and GlcNAc-transferase activity with Mn2+, respectively, 0.2 mM Pasteurella multocida
Mn2+ absolutely required, best metal cofactor, 0.2 mM Pasteurella multocida
additional information metal requirements of mutant enzymes, overview Pasteurella multocida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Pasteurella multocida the produced hyaluronan capsule enhances infection ?
-
?
additional information Pasteurella multocida type A the produced hyaluronan capsule enhances infection ?
-
?

Organism

Organism UniProt Comment Textmining
Pasteurella multocida
-
type A
-
Pasteurella multocida type A
-
type A
-

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] 2 active sites, the 2 DXD motifs of the transferase sites in the C-terminus, residues 686-703, are essential for activity Pasteurella multocida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the produced hyaluronan capsule enhances infection Pasteurella multocida ?
-
?
additional information substrate specificity, the GlcNAc-transferase, but not the GlcUA-transferase activity depends on the WGGED motif, overview Pasteurella multocida ?
-
?
additional information the produced hyaluronan capsule enhances infection Pasteurella multocida type A ?
-
?
additional information substrate specificity, the GlcNAc-transferase, but not the GlcUA-transferase activity depends on the WGGED motif, overview Pasteurella multocida type A ?
-
?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate formation of linear hyaluronan polymers composed of alternating beta3-N-acetylglucosamine-beta4-glucuronic acid Pasteurella multocida [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP the product chain length can grow at the reducing end up to 40000 monosaccharides with a MW of over 8 million Da before it is released by the class I enzyme ?
UDP-N-acetyl-D-glucosamine + UDP-D-glucuronate formation of linear hyaluronan polymers composed of alternating beta3-N-acetylglucosamine-beta4-glucuronic acid Pasteurella multocida type A [beta-N-acetyl-D-glucosaminyl(1-4)beta-D-glucuronosyl(1-3)]n + UDP the product chain length can grow at the reducing end up to 40000 monosaccharides with a MW of over 8 million Da before it is released by the class I enzyme ?

Synonyms

Synonyms Comment Organism
HA synthase
-
Pasteurella multocida
HAS
-
Pasteurella multocida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Pasteurella multocida