Cloned (Comment) | Organism |
---|---|
gene hyaD, recombinant expression of the soluble C-terminally His6-tagged PmHAS1-703 truncated enzyme from pET101/D-TOPO expression vector with an additional V5 epitope | Pasteurella multocida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
UMP | dead-end inhibitor | Pasteurella multocida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | structural and kinetic analysis and modeling, both NAc- and UA-transferase domains follow a sequential kinetic mechanism, most likely an ordered one in which the UDP-sugar donor binds first, followed by the HA oligosaccharide. After transfer of the sugar moiety, both products are released, first the elongated HA oligosaccharide and then the UDP sugar. A mechanistic shift from a steady-state ordered bi-bi to rapid equilibrium ordered bi-bi mechanism is observed at the NAc-site between the HA6 and HA8 elongation, detailed overview | Pasteurella multocida | |
0.6 | - |
hyaluronan oligomer HA5 | with UDP-alpha-D-glucuronate, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida | |
0.7 | - |
hyaluronan oligomer HA4 | with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida | |
0.7 | - |
hyaluronan oligomer HA8 | with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida | |
1 | - |
hyaluronan oligomer HA6 | with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Pasteurella multocida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Pasteurella multocida | Pasteurella multocida hyaluronan synthase encompasses two transferase domains that elongate a growing hyaluronan oligosaccharide chain by addition of either GlcNAc or GlcUA residues from a corresponding UDP-sugar | ? | - |
? | |
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] | Pasteurella multocida | - |
UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] | - |
? | |
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] | Pasteurella multocida | - |
UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pasteurella multocida | Q7BLV3 | gene hyaD | - |
Purification (Comment) | Organism |
---|---|
recombinant soluble C-terminally His6-tagged PmHAS1-703 by nickel affinity chromatography | Pasteurella multocida |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] = UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] | a mechanistic shift from a steady-state ordered bi-bi to rapid equilibrium ordered bi-bi mechanism is observed at the NAc-site between the HA6 and HA8 elongation | Pasteurella multocida | |
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] | a mechanistic shift from a steady-state ordered bi-bi to rapid equilibrium ordered bi-bi mechanism is observed at the NAc-site between the HA6 and HA8 elongation | Pasteurella multocida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Pasteurella multocida hyaluronan synthase encompasses two transferase domains that elongate a growing hyaluronan oligosaccharide chain by addition of either GlcNAc or GlcUA residues from a corresponding UDP-sugar | Pasteurella multocida | ? | - |
? | |
additional information | initial velocity studies of single-step elongations are conducted for both domains by independently varying the concentrations of the hyaluronan oligosaccharide and the UDP-sugar. Two-substrate models are discriminated by their goodness-of-fit parameters and by dead-end inhibition studies. Coupled-enzyme assay using LDH, PK; NADH and phosphoenolpyruvate, as well as hyaluronan oligosaccharides, UDP-GlcNAc and UDP-GlcUA, overview | Pasteurella multocida | ? | - |
? | |
UDP-alpha-D-glucuronate + hyaluronan oligomer HA5 | - |
Pasteurella multocida | UDP + beta-D-glucuronosyl-(1->3)-hyaluronan oligomer HA5 | - |
? | |
UDP-alpha-D-glucuronate + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] | - |
Pasteurella multocida | UDP + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-[nascent hyaluronan] | - |
? | |
UDP-alpha-N-acetyl-D-glucosamine + beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] | - |
Pasteurella multocida | UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-beta-D-glucuronosyl-(1->3)-N-acetyl-beta-D-glucosaminyl-(1->4)-[nascent hyaluronan] | - |
? | |
UDP-alpha-N-acetyl-D-glucosamine + hyaluronan oligomer HA4 | - |
Pasteurella multocida | UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-hyaluronan oligomer HA4 | - |
? | |
UDP-alpha-N-acetyl-D-glucosamine + hyaluronan oligomer HA6 | - |
Pasteurella multocida | UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-hyaluronan oligomer HA6 | - |
? | |
UDP-alpha-N-acetyl-D-glucosamine + hyaluronan oligomer HA8 | - |
Pasteurella multocida | UDP + N-acetyl-beta-D-glucosaminyl-(1->4)-hyaluronan oligomer HA8 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PmHAS | - |
Pasteurella multocida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
assay at | Pasteurella multocida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.9 | - |
hyaluronan oligomer HA5 | with UDP-alpha-D-glucuronate, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida | |
7.9 | - |
hyaluronan oligomer HA4 | with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida | |
8.8 | - |
hyaluronan oligomer HA8 | with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida | |
13.5 | - |
hyaluronan oligomer HA6 | with UDP-alpha-N-acetyl-D-glucosamine, pH 8.0, 35°C, recombinant enzyme | Pasteurella multocida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pasteurella multocida |