A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P)

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Standard

A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P). / Hingst, Janne Rasmuss; Bjerre, Rie Dybboe; Wojtaszewski, Jørgen; Jensen, Jørgen.

I: Journal of Applied Physiology, 11.07.2019.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Hingst, JR, Bjerre, RD, Wojtaszewski, J & Jensen, J 2019, 'A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P)', Journal of Applied Physiology. https://doi.org/10.1152/japplphysiol.00207.2018

APA

Hingst, J. R., Bjerre, R. D., Wojtaszewski, J., & Jensen, J. (Accepteret/In press). A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P). Journal of Applied Physiology. https://doi.org/10.1152/japplphysiol.00207.2018

Vancouver

Hingst JR, Bjerre RD, Wojtaszewski J, Jensen J. A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P). Journal of Applied Physiology. 2019 jul 11. https://doi.org/10.1152/japplphysiol.00207.2018

Author

Hingst, Janne Rasmuss ; Bjerre, Rie Dybboe ; Wojtaszewski, Jørgen ; Jensen, Jørgen. / A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P). I: Journal of Applied Physiology. 2019.

Bibtex

@article{7ebc06e3b1d9483d9b6e9293223172bc,
title = "A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P)",
abstract = "Glucose phosphorylation by hexokinase (HK) is a rate-limiting step in glucose metabolism. Regulation of HK includes feedback inhibition by its product glucose-6-phosphate (G6P) and mitochondria binding. HK affinity for G6P is difficult to measure because its natural product (G6P) inhibits enzyme activity. HK phosphorylates several hexoses and we have taken advantage of the fact that 2-deoxyglucose-6-phosphate (2-DG-6P) does not inhibit HK activity. By this we have developed a new method for rapid radiochemical analysis of HK activity with 2-deoxyglucose (2-DG) as substrate, which allows control of the concentrations of G6P to investigate HK affinity for inhibition by G6P. We verified that 2-DG serves as a substrate for the HK reaction with linear time and concentration dependency as well as expected Vmax and KM. This is the first simple assay evaluating feedback inhibition of HK by its product G6P and provides a unique technique for future research evaluating the regulation of glucose phosphorylation under various physiological conditions.",
author = "Hingst, {Janne Rasmuss} and Bjerre, {Rie Dybboe} and J{\o}rgen Wojtaszewski and J{\o}rgen Jensen",
note = "Afventer publicering som [Epub ahead of print] samt til deling af CURIS-nummer.",
year = "2019",
month = "7",
day = "11",
doi = "10.1152/japplphysiol.00207.2018",
language = "English",
journal = "Journal of Applied Physiology",
issn = "8750-7587",
publisher = "American Physiological Society",

}

RIS

TY - JOUR

T1 - A rapid radiochemical filter paper assay for determination of Hexokinase activity and affinity for glucose-6-phosphate (G6P)

AU - Hingst, Janne Rasmuss

AU - Bjerre, Rie Dybboe

AU - Wojtaszewski, Jørgen

AU - Jensen, Jørgen

N1 - Afventer publicering som [Epub ahead of print] samt til deling af CURIS-nummer.

PY - 2019/7/11

Y1 - 2019/7/11

N2 - Glucose phosphorylation by hexokinase (HK) is a rate-limiting step in glucose metabolism. Regulation of HK includes feedback inhibition by its product glucose-6-phosphate (G6P) and mitochondria binding. HK affinity for G6P is difficult to measure because its natural product (G6P) inhibits enzyme activity. HK phosphorylates several hexoses and we have taken advantage of the fact that 2-deoxyglucose-6-phosphate (2-DG-6P) does not inhibit HK activity. By this we have developed a new method for rapid radiochemical analysis of HK activity with 2-deoxyglucose (2-DG) as substrate, which allows control of the concentrations of G6P to investigate HK affinity for inhibition by G6P. We verified that 2-DG serves as a substrate for the HK reaction with linear time and concentration dependency as well as expected Vmax and KM. This is the first simple assay evaluating feedback inhibition of HK by its product G6P and provides a unique technique for future research evaluating the regulation of glucose phosphorylation under various physiological conditions.

AB - Glucose phosphorylation by hexokinase (HK) is a rate-limiting step in glucose metabolism. Regulation of HK includes feedback inhibition by its product glucose-6-phosphate (G6P) and mitochondria binding. HK affinity for G6P is difficult to measure because its natural product (G6P) inhibits enzyme activity. HK phosphorylates several hexoses and we have taken advantage of the fact that 2-deoxyglucose-6-phosphate (2-DG-6P) does not inhibit HK activity. By this we have developed a new method for rapid radiochemical analysis of HK activity with 2-deoxyglucose (2-DG) as substrate, which allows control of the concentrations of G6P to investigate HK affinity for inhibition by G6P. We verified that 2-DG serves as a substrate for the HK reaction with linear time and concentration dependency as well as expected Vmax and KM. This is the first simple assay evaluating feedback inhibition of HK by its product G6P and provides a unique technique for future research evaluating the regulation of glucose phosphorylation under various physiological conditions.

U2 - 10.1152/japplphysiol.00207.2018

DO - 10.1152/japplphysiol.00207.2018

M3 - Journal article

JO - Journal of Applied Physiology

JF - Journal of Applied Physiology

SN - 8750-7587

ER -

ID: 224997908