Studies on the mechanism of action of the histone kinase dependent on adenosine 3',5'-monophosphate. Investigation of protein-protein interaction by electron spin-resonance spectroscopy and stopped-flow methods.
The interaction of the cAMP-dependent protein kinase catalytic subunit with its protein substrate, histone H1, was studied. The H1 molecule was specifically converted into the aminotyrosine-72 derivative. Fluorescent and spin labels were introduced into this residue. The changes in the ESR and fluorescence spectra of respective derivatives were observed upon the interaction of the latter with the catalytic subunit, thus enabling us to determine some kinetic and equilibrium parameters of this process. Stopped-flow investigation of the transient phase of the binding reaction indicates that the kinetic curve is described by a three-exponential function. The rate of protein-protein interaction is close to the rate of phosphate transfer from phosphoenzyme intermediate to protein substrate.
D000818:Animals; D001921:Brain / Q000201:enzymology; D002384:Catalysis; D055598:Chemical Phenomena; D002621:Chemistry; D000242:Cyclic AMP / Q000494:pharmacology*; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D006656:Protamine Kinase / Q000502:physiology*; D011494:Protein Kinases / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D013552:Swine
D000242:Cyclic AMP; D011494:Protein Kinases; D006656:Protamine Kinase