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Literature summary for 2.4.3.1 extracted from

  • Meng, L.; Forouhar, F.; Thieker, D.; Gao, Z.; Ramiah, A.; Moniz, H.; Xiang, Y.; Seetharaman, J.; Milaninia, S.; Su, M.; Bridger, R.; Veillon, L.; Azadi, P.; Kornhaber, G.; Wells, L.; Montelione, G.T.; Woods, R.J.; Tong, L.; Moremen, K.W.
    Enzymatic basis for N-glycan sialylation: structure of rat alpha2,6-sialyltransferase (ST6GAL1) reveals conserved and unique features for glycan sialylation (2013), J. Biol. Chem., 288, 34680-34698.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.4.3.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in HEK-293 and 293-F cells Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment) Organism
microbatch method, using 100 mM Tris, pH 9, 40% (w/v) PEG 1000, and 100 mM lithium bromide Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
C350A/C361A inactive Rattus norvegicus
D271A the mutant shows strongly reduced activity compared to the wild type enzyme Rattus norvegicus
F208A the mutant shows strongly reduced activity compared to the wild type enzyme Rattus norvegicus
H367A inactive Rattus norvegicus
K355A the mutant shows increased activity compared to the wild type enzyme Rattus norvegicus
N230A inactive Rattus norvegicus
Q232A the mutant shows strongly reduced activity compared to the wild type enzyme Rattus norvegicus
Q354A the mutant shows reduced activity compared to the wild type enzyme Rattus norvegicus
Y119F the mutant shows reduced activity compared to the wild type enzyme Rattus norvegicus
Y272A the mutant shows strongly reduced activity compared to the wild type enzyme Rattus norvegicus
Y366A inactive Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.078
-
 CMP-N-acetylneuraminate mutant enzyme Y119F, at pH 6.5 and 37°C Rattus norvegicus
0.092
-
 CMP-N-acetylneuraminate recombinant wild type enzyme from 293-F cells, at pH 6.5 and 37°C Rattus norvegicus
0.093
-
 CMP-N-acetylneuraminate recombinant wild type enzyme from HEK-293 cells, at pH 6.5 and 37°C Rattus norvegicus
0.095
-
 CMP-N-acetylneuraminate mutant enzyme K355A, at pH 6.5 and 37°C Rattus norvegicus
0.1
-
 CMP-N-acetylneuraminate mutant enzyme Q354A, at pH 6.5 and 37°C Rattus norvegicus
0.133
-
 CMP-N-acetylneuraminate mutant enzyme Q232A, at pH 6.5 and 37°C Rattus norvegicus
0.22
-
 CMP-N-acetylneuraminate mutant enzyme Y272A, at pH 6.5 and 37°C Rattus norvegicus
0.365
-
 CMP-N-acetylneuraminate mutant enzyme D271A, at pH 6.5 and 37°C Rattus norvegicus
0.4
-
 CMP-N-acetylneuraminate mutant enzyme F208A, at pH 6.5 and 37°C Rattus norvegicus
1
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme F208A, at pH 6.5 and 37°C Rattus norvegicus
1.1
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme K355A, at pH 6.5 and 37°C Rattus norvegicus
1.4
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine recombinant wild type enzyme from HEK-293 cells, at pH 6.5 and 37°C Rattus norvegicus
1.8
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Q354A, at pH 6.5 and 37°C Rattus norvegicus
1.8
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Y119F, at pH 6.5 and 37°C Rattus norvegicus
1.8
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine recombinant wild type enzyme from 293-F cells, at pH 6.5 and 37°C Rattus norvegicus
2.6
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Q232A, at pH 6.5 and 37°C Rattus norvegicus
3.5
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Y272A, at pH 6.5 and 37°C Rattus norvegicus
6.5
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme D271A, at pH 6.5 and 37°C Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
CMP-N-acetylneuraminate + beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine Rattus norvegicus
-
 CMP + alpha-N-acetylneuraminyl-(2->6)-beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine
-
 ?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P13721
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Superdex-75 gel filtration Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CMP-N-acetylneuraminate + beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine
-
 Rattus norvegicus CMP + alpha-N-acetylneuraminyl-(2->6)-beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine
-
 ?

Synonyms

Synonyms Comment Organism
alpha2,6-sialyltransferase
-
 Rattus norvegicus
ST6Gal1
-
 Rattus norvegicus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.05
-
 CMP-N-acetylneuraminate mutant enzyme D271A, at pH 6.5 and 37°C Rattus norvegicus
0.05
-
 CMP-N-acetylneuraminate mutant enzyme Y272A, at pH 6.5 and 37°C Rattus norvegicus
0.07
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Y272A, at pH 6.5 and 37°C Rattus norvegicus
0.09
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme D271A, at pH 6.5 and 37°C Rattus norvegicus
0.1
-
 CMP-N-acetylneuraminate mutant enzyme Q232A, at pH 6.5 and 37°C Rattus norvegicus
0.14
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Q232A, at pH 6.5 and 37°C Rattus norvegicus
0.19
-
 CMP-N-acetylneuraminate mutant enzyme F208A, at pH 6.5 and 37°C Rattus norvegicus
0.31
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Y119F, at pH 6.5 and 37°C Rattus norvegicus
0.35
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine recombinant wild type enzyme from HEK-293 cells, at pH 6.5 and 37°C Rattus norvegicus
0.39
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme K355A, at pH 6.5 and 37°C Rattus norvegicus
0.39
-
 CMP-N-acetylneuraminate mutant enzyme Y119F, at pH 6.5 and 37°C Rattus norvegicus
0.42
-
 CMP-N-acetylneuraminate mutant enzyme Q354A, at pH 6.5 and 37°C Rattus norvegicus
0.43
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme F208A, at pH 6.5 and 37°C Rattus norvegicus
0.49
-
 CMP-N-acetylneuraminate recombinant wild type enzyme from HEK-293 cells, at pH 6.5 and 37°C Rattus norvegicus
0.59
-
 CMP-N-acetylneuraminate recombinant wild type enzyme from 293-F cells, at pH 6.5 and 37°C Rattus norvegicus
0.64
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Q354A, at pH 6.5 and 37°C Rattus norvegicus
0.67
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine recombinant wild type enzyme from 293-F cells, at pH 6.5 and 37°C Rattus norvegicus
0.69
-
 CMP-N-acetylneuraminate mutant enzyme K355A, at pH 6.5 and 37°C Rattus norvegicus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.01
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme D271A, at pH 6.5 and 37°C Rattus norvegicus
0.02
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Y272A, at pH 6.5 and 37°C Rattus norvegicus
0.05
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Q232A, at pH 6.5 and 37°C Rattus norvegicus
0.13
-
 CMP-N-acetylneuraminate mutant enzyme D271A, at pH 6.5 and 37°C Rattus norvegicus
0.17
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Y119F, at pH 6.5 and 37°C Rattus norvegicus
0.23
-
 CMP-N-acetylneuraminate mutant enzyme Y272A, at pH 6.5 and 37°C Rattus norvegicus
0.25
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine recombinant wild type enzyme from HEK-293 cells, at pH 6.5 and 37°C Rattus norvegicus
0.36
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme K355A, at pH 6.5 and 37°C Rattus norvegicus
0.36
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme Q354A, at pH 6.5 and 37°C Rattus norvegicus
0.37
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine recombinant wild type enzyme from 293-F cells, at pH 6.5 and 37°C Rattus norvegicus
0.44
-
 beta-D-galactosyl-(1->4)-N-acetyl-beta-D-glucosamine mutant enzyme F208A, at pH 6.5 and 37°C Rattus norvegicus
0.47
-
 CMP-N-acetylneuraminate mutant enzyme F208A, at pH 6.5 and 37°C Rattus norvegicus
4.17
-
 CMP-N-acetylneuraminate mutant enzyme Q354A, at pH 6.5 and 37°C Rattus norvegicus
5
-
 CMP-N-acetylneuraminate mutant enzyme Y119F, at pH 6.5 and 37°C Rattus norvegicus
5.27
-
 CMP-N-acetylneuraminate recombinant wild type enzyme from HEK-293 cells, at pH 6.5 and 37°C Rattus norvegicus
6.45
-
 CMP-N-acetylneuraminate recombinant wild type enzyme from 293-F cells, at pH 6.5 and 37°C Rattus norvegicus
7.3
-
 CMP-N-acetylneuraminate mutant enzyme K355A, at pH 6.5 and 37°C Rattus norvegicus