Localization of the High‐Affinity ATP Site in Adenosine‐3′:5′‐monophosphate‐Dependent Protein Kinase Type I
Open Access
- 1 January 1979
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 93 (1), 141-146
- https://doi.org/10.1111/j.1432-1033.1979.tb12804.x
Abstract
8-Azido-adenosine 5''-triphosphate .**GRAPHIC**. appeared to be a suitable photoaffinity label for c[cyclic]AMP dependent protein kinase [EC 2.7.1.37]. It competed with ATP for the high-affinity ATP site in the undissociated form of the kinase and in the phosphotransferase reaction catalyzed by the catalytic subunit. It is accepted as a substrate in the phosphotransfer reaction. .**GRAPHIC**. incorporated into the holoenzyme is covalently bound upon irradiation. Protection experiments with ATP indicated that this covalent attachment occurs in the high-affinity ATP site of the enzyme. Polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate shows that .**GRAPHIC**. is bound to the catalytic subunit. After irradiation the enzyme was dissociated by cAMP. Proportional to the incorporated .**GRAPHIC**. a loss in phosphotransferase activity was found. Both ATP sites probably coincide with respect to their adenine binding part. Thus binding of the regulatory subunit to the catalytic subunit would then transform the low-affinity catalytically active ATP site into a high-affinity inactive site.This publication has 31 references indexed in Scilit:
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