The inhibitory mechanism of in vitro protein phosphorylation by a nonprotein chromophore removed from neocarzinostatin.
The inhibitory effect of a nonprotein chromophore removed from neocarzinostatin on protein phosphorylation by nuclear protein kinase in vitro has been studied. Low levels of the chromophore greatly inhibited protein phosphorylation in vitro. This inhibition, however, was not selectively dependent on the indicated kinases and their different phosphate acceptors (histones and non-histone protein). In contrast, the protein component (apoprotein) of neocarzinostatin did not affect the phosphorylation even at a concentration of 400-times higher than that of the chromophore. Moreover, apoprotein suppressed the chromophore-induced inhibition of protein phosphorylation in vitro in proportion to the apoprotein concentrations. Kinetic and analytical experiments suggest that the chromophore-induced inhibition of protein phosphorylation seems to be due to the binding of the chromophore to the kinases. In addition, we found that ultraviolet irradiation as well as methanol extraction can release the chromophore from neocarzinostatin, but it exhibits no inhibitory activity of DNA synthesis in growing cells. The fact that the chromophore-induced inhibition of protein phosphorylation in vitro was not sensitive to ultraviolet irradiation, which rapidly inactivated the ability of the chromophore to induce DNA degradation in vitro, suggests that there are different actions involved in the two inhibitions induced by the chromophore which is removed from neocarzinostatin.
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000302:isolation & purification*; D002467:Cell Nucleus / Q000201:enzymology; D000242:Cyclic AMP / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D010766:Phosphorylation; D006656:Protamine Kinase / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D013154:Spleen / Q000201:enzymology; D013379:Substrate Specificity; D009353:Zinostatin / Q000302:isolation & purification*
D000903:Antibiotics, Antineoplastic; D009353:Zinostatin; D000242:Cyclic AMP; D011494:Protein Kinases; D006656:Protamine Kinase