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Kinetic analysis of gluconate phosphorylation by human gluconokinase using isothermal titration calorimetry
Author(s) -
Rohatgi Neha,
Guðmundsson Steinn,
Rolfsson Óttar
Publication year - 2015
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2015.10.024
Subject(s) - isothermal titration calorimetry , calorimetry , chemistry , ternary complex , titration , enzyme , isothermal process , substrate (aquarium) , ternary operation , kinetics , catalysis , reaction mechanism , biochemistry , inorganic chemistry , thermodynamics , biology , ecology , physics , quantum mechanics , programming language , computer science
Gluconate is a commonly encountered nutrient, which is degraded by the enzyme gluconokinase to generate 6‐phosphogluconate. Here we used isothermal titration calorimetry to study the properties of this reaction. Δ H , K M and k cat are reported along with substrate binding data. We propose that the reaction follows a ternary complex mechanism, with ATP binding first. The reaction is inhibited by gluconate, as it binds to an Enzyme–ADP complex forming a dead‐end complex. The study exemplifies that ITC can be used to determine mechanisms of enzyme catalyzed reactions, for which it is currently not commonly applied.