The Isolation of a Cyclic Nucleotide Independent, Trypsin Sensitive Proenzyme and Its Activation of Tyrosine Hydroxylase
Stier, Michael A.
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It will be shown that a cyclic nucleotide independent, trypsin sensitive protein kinase can be isolated and purified from whole rat brain homogenate. The purification utilizes several common biochemical separation techniques: anion exchange chromatography; Sephadex gel chromatography and Isoelectric focusing electrophoresis (I.E.F.). The proenzyme which was isolated from rat brain was found to phosphoryllate, Histone type II B in the presence of 2-mercaptoethanol and Mg2+ • A .4 pm concentration of trypsin activated. the protein kinase's phosphoryllation of histone as substrate in the standard kinase assay. A calcium ion sensitivity has also been observed in the proenzyme. The purified proenzyme is a soluble and acidic protein which will be explored in several ways. The purified proenzyme was examined to see if it could activate other enzyme systems. We will show that the addition of our purified cyclic nucleotide independent protein kinase, to a system containing its optimal phosphoryllating conditions and adrenal homogenate causes an activation of TOH.