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Literature summary for 3.2.1.18 extracted from

  • Damager, I.; Buchini, S.; Amaya, M.F.; Buschiazzo, A.; Alzari, P.; Frasch, A.C.; Watts, A.; Withers, S.G.
    Kinetic and mechanistic analysis of Trypanosoma cruzi trans-sialidase reveals a classical ping-pong mechanism with acid/base catalysis (2008), Biochemistry, 47, 3507-3512.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
D59A replacement of putative acid/base catalyst, demonstration of the half-reaction with formation of sialyl-enzyme intermediate. Activity is restored by addition of azide and a sialyl azide product is formed Trypanosoma cruzi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
4-nitrophenyl alpha-sialoside mutant D59A, hydrolysis, pH 7.6, 25°C Trypanosoma cruzi
0.06
-
(trifluoromethyl)umbelliferyl alpha-sialoside wild-type, hydrolysis, pH 7.6, 25°C Trypanosoma cruzi
1.09
-
4-nitrophenyl alpha-sialoside wild-type, hydrolysis, pH 7.6, 25°C Trypanosoma cruzi

Organism

Organism UniProt Comment Textmining
Trypanosoma cruzi
-
-
-

Reaction

Reaction Comment Organism Reaction ID
colominic acid + H2O = sialic acid + lactose ping-pong bi-bi kinetic mechanism. Residue D59 functions as acid/base catalyst, and Y342 as active site nucleophile Trypanosoma cruzi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(trifluoromethyl)umbelliferyl alpha-sialoside + H2O
-
Trypanosoma cruzi (trifluoromethyl)umbelliferone + sialic acid
-
?
(trifluoromethyl)umbelliferyl alpha-sialoside + lactose
-
Trypanosoma cruzi (trifluoromethyl)umbelliferone + sialyllactose
-
?
4-nitrophenyl alpha-sialoside + H2O
-
Trypanosoma cruzi 4-nitrophenol + sialic acid
-
?
4-nitrophenyl alpha-sialoside + lactose
-
Trypanosoma cruzi 4-nitrophenol + sialyllactose
-
?
additional information in addition to hydrolysis, enzyme catalyzes transglycosylation of sialyl residues to lactose. Km value and kcat value for sialyl transfer from 4-nitrophenyl alpha-sialoside are 5.8 mM and 6.77 per s, for sialyl transfer from (trifluoromethyl)umbelliferyl alpha-sialoside 0.26 mM and 15.7 per s, respectively Trypanosoma cruzi ?
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.031
-
4-nitrophenyl alpha-sialoside mutant D59A, hydrolysis, pH 7.6, 25°C Trypanosoma cruzi
1.57
-
4-nitrophenyl alpha-sialoside wild-type, hydrolysis, pH 7.6, 25°C Trypanosoma cruzi
2.9
-
(trifluoromethyl)umbelliferyl alpha-sialoside wild-type, hydrolysis, pH 7.6, 25°C Trypanosoma cruzi