Affinity labeling of adenine nucleotide-related enzymes with reactive adenine nucleotide analogs. I. Affinity labeling of glyceraldehyde 3-phosphate dehydrogenase and myokinase with a reactive AMP analog.:I. Affinity Labeling of Glyceraldehyde 3-Phosphate Dehydrogenase and Myokinase with a Reactive AMP Analog

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  • SUZUKI Koichi
    Department of Biochemistry, Faculty of Medicine, the University of Tokyo
  • EGUCHI Chikahiko
    Department of Biochemistry, Faculty of Medicine, the University of Tokyo
  • IMAHORI Kazutomo
    Department of Biochemistry, Faculty of Medicine, the University of Tokyo

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Description

Rabbit muscle glyceraldehyde 3-phosphate dehydrogenase (GPD) and myokinase (MK) were rapidly inactivated by a reactive AMP analog, N6-(p-bromoacetaminobenzyl)-AMP, under mild conditions. Complete inactivation was observed when 4 and 0.3 mol of the reagent with respect to enzyme were reacted with GPD and MK, respectively. The inactivation of both enzymes was favored at higher pH and the enzymes were protected by addition of adenine nucleotide substrate. Modified GPD or MK had no affinity for AMP-Sepharose, in contrast to the native enzymes. From these results, the inactivation of GPD and MK by the reactive AMP analog can be regarded as an affinity labeling. The possibility that the present AMP analog may be used as a general affinity labeling reagent for various adenine nucleotide-related enzymes is discussed based on the results obtained.

Journal

  • J Biochem (Tokyo)

    J Biochem (Tokyo) 81 (4), 1147-1154, 1977

    The Japanese Biochemical Society

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