Method for measuring glucosamine-6-phosphate
The present invention pertains to methods for measuring the activity of glutamine:fructose 6-phosphate amidotransferase which comprise the steps of (a) forming a mixture of glutamine:fructose 6-phosphate amidotransferase, fructose 6-phosphate, and glutamine; (b) incubating the mixture under physiolo...
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Sprache: | eng ; fre ; ger |
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Zusammenfassung: | The present invention pertains to methods for measuring the activity of glutamine:fructose 6-phosphate amidotransferase which comprise the steps of (a) forming a mixture of glutamine:fructose 6-phosphate amidotransferase, fructose 6-phosphate, and glutamine; (b) incubating the mixture under physiological conditions to allow the formation of glucosamine 6-phosphate; (c) acetylating the glucosamine 6-phosphate to form N-acetylglucosamine 6-phosphate; (d) reacting the N-acetylglucosamine 6-phosphate with Ehrlich's reagent; and (e) measuring the amount of N-acetylglucosamine 6-phosphate by determining the optical density at 500-610nm of the mixture and comparing the amount of N-acetylglucosamine 6-phosphate with control samples. The present invention also pertains to methods for measuring the inhibitory activity of a test compound on glutamine:fructose 6-phosphate amidotransferase, which comprise the steps of (a) forming a mixture of a test compound, glutamine:fructose 6-phosphate amidotransferase, fructose 6-phosphate, and glutamine; (b) incubating the mixture under physiological conditions to allow the test compound to inhibit glutamine:fructose 6-phosphate amidotransferase from forming glucosamine 6-phosphate; (c) acetylating the glucosamine 6-phosphate to form N-acetylglucosamine 6-phosphate; (d) reacting the N-acetylglucosamine 6-phosphate with Ehrlich's reagent; and (e) measuring the amount of N-acetylglucosamine 6-phosphate by determining the optical density at 500-610nm of the mixture and comparing the amount of N-acetylglucosamine 6-phosphate with control samples. |
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