Kinetic analysis of gluconate phosphorylation by human gluconokinase using isothermal titration calorimetry

Neha Rohatgi; Steinn Gudmundsson; Óttar Rolfsson

doi:10.1016/j.febslet.2015.10.024

Kinetic analysis of gluconate phosphorylation by human gluconokinase using isothermal titration calorimetry

Neha Rohatgi, Steinn Gudmundsson, Óttar Rolfsson

University of Iceland

Research output: Contribution to journal › Article › peer-review

Abstract

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. ΔH, 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.

Original language	English
Pages (from-to)	3548-3555
Number of pages	8
Journal	FEBS Letters
Volume	589
Issue number	23
DOIs	https://doi.org/10.1016/j.febslet.2015.10.024
Publication status	Published - 30 Nov 2015

Bibliographical note

Other keywords

Enzyme kinetics
Gluconate (Glcn)
Human gluconokinase (hGntK, IdnK)
Human metabolism
Isothermal titration calorimetry

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10.1016/j.febslet.2015.10.024

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title = "Kinetic analysis of gluconate phosphorylation by human gluconokinase using isothermal titration calorimetry",

abstract = "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. ΔH, KM and kcat 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.",

keywords = "Enzyme kinetics, Gluconate (Glcn), Human gluconokinase (hGntK, IdnK), Human metabolism, Isothermal titration calorimetry",

author = "Neha Rohatgi and Steinn Gudmundsson and {\'O}ttar Rolfsson",

note = "Publisher Copyright: {\textcopyright} 2015 Federation of European Biochemical Societies.",

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doi = "10.1016/j.febslet.2015.10.024",

language = "English",

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T1 - Kinetic analysis of gluconate phosphorylation by human gluconokinase using isothermal titration calorimetry

AU - Rohatgi, Neha

AU - Gudmundsson, Steinn

AU - Rolfsson, Óttar

PY - 2015/11/30

Y1 - 2015/11/30

N2 - 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. ΔH, KM and kcat 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.

AB - 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. ΔH, KM and kcat 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.

KW - Enzyme kinetics

KW - Gluconate (Glcn)

KW - Human gluconokinase (hGntK, IdnK)

KW - Human metabolism

KW - Isothermal titration calorimetry

UR - https://www.scopus.com/pages/publications/84947284014

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DO - 10.1016/j.febslet.2015.10.024

M3 - Article

C2 - 26505675

SN - 0014-5793

VL - 589

SP - 3548

EP - 3555

JO - FEBS Letters

JF - FEBS Letters

IS - 23

ER -

Kinetic analysis of gluconate phosphorylation by human gluconokinase using isothermal titration calorimetry

Abstract

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