Regulation of cyclic nucleotide-dependent protein kinase activity by histones and poly(L-arginine).
Interaction of cGMP-dependent protein kinase with histones H2A, H2B, H3, and H4, or poly(L-arginine) resulted in changes in enzyme conformation such that inactivation of cGMP binding and activation of basal catalytic activity (assayed without cGMP) occurred. Total kinase activity as determined by phosphorylation of exogenous substrates subsequently decreased, but autophosphorylation of the enzyme was enhanced. The reaction was specific for nucleosome core histones and poly(L-arginine); H1, troponin, and poly(L-lysine) had no effect. Inactivation of cyclic nucleotide binding sites followed pseudo-first order kinetics and, at various histone concentrations, exhibited saturation kinetics at low ionic strength (2 mM potassium phosphate, pH 6.8), but non-saturation kinetics at higher ionic strength (37.5 mM potassium phosphate, pH 6.8, 12.5 mM MgCl2). Saturation kinetics was observed with poly(L-arginine) at both low and high ionic strength. Kinetic parameters measured under saturation conditions were determined for each core histone and poly(L-arginine). Core histones and poly(L-arginine) were noncompetitive inhibitors of cGMP binding; core histones and poly(L-arginine) interacted competitively at an enzyme site designated as the poly(L-arginine) binding site. Regulatory subunits of cAMP-dependent protein kinase contain a similar poly(L-arginine) binding site. Modulator proteins bind to poly(L-arginine) or arginyl residues in histone to prevent interaction with the poly(L-arginine) binding site on the enzymes. Through this mechanism, modulator proteins maintain cyclic nucleotide dependency and full enzyme activity.
D000818:Animals; D002417:Cattle; D006152:Cyclic GMP / Q000494:pharmacology; D006657:Histones / Q000494:pharmacology*; D007700:Kinetics; D008168:Lung / Q000201:enzymology; D010455:Peptides / Q000494:pharmacology*; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013950:Thymus Gland
D006657:Histones; D010455:Peptides; C015462:polyarginine; D011494:Protein Kinases; D006152:Cyclic GMP