Hagiwara Masatoshi

Differential effects of flavonoids as inhibitors of tyrosine protein kinases and serine/threonine protein kinases.

Abstract The inhibitory potencies of bioflavonoids on various tyrosine protein kinases and serine/ threonine protein kinases were investigated. The phosphotransferase activity of an oncogene product, pp130fps, and a growth factor receptor, insulin receptor, were inhibited by myricetin, a derivative of quercetin, However, tyrosine kinase activity in the particulate fraction from human platelets (PM-TPK) was resistant to myricetin. Apparent Ki values of myricetin for tyrosine protein kinases of pp130fps and insulin receptor were 1.8 and 2.6 μM, respectively. The Ki values for serine/threonine kinase activities of myosin light chain kinase (MLC-kinase), casein kinase I, casein kinase II, cAMP-dependent protein kinase, and protein kinase C were 1.7,μM, 9.0 μM, 0.6 μM, 27.5 μM, and 12.1 μM, respectively. Lineweaver-Burk plots revealed that myricetin competitively inhibits pp130fps tyrosine kinase, myosin light chain kinase, casein kinase I and II with ATP, but does not inhibit other protein kinases. Since myricetin is a hydroxylated derivative of quercetin, the inhibitory effects of a series of seven flavonoids with various numbers of hydroxy residues were examined. Structure activity studies exhibited that the inhibitory potencies of the flavonoids for tyrosine kinases of pp130fps and insulin receptor correlated with the number of hydroxy residues on the flavone rings (γ = 0.974 and 0.926, respectively), whereas the hydroxylation influenced to a lesser extent the inhibitory potencies for serine/threonine protein kinase. The hydroxy residues at position 3 and 5 did not affect the activities of cAMP-dependent protein kinase, and protein kinase C, and the hydroxylation at position 5 is detrimental for the inhibition of MLC-kinase, and casein kinase I and II. Thus, flavonoids may be useful tools to elucidate the active site of tyrosine and serine/threonine protein kinases.