Abstract
A synthetic peptide derived from the phosphorylation site in the β-subunit of phosphorylase kinase (RTKRSGSVYEPLKI) is an efficient substrate for rat brain protein kinase C: K m = 18±2 μM and V max = 2.1±0.1 μmol/min/mg. The phosphorylation of the peptide, which occurs at Ser 7, can be followed by four independent procedures. 1. Standard measurement of 32P incorporation. 2. Reverse phase HPLC in a gradient system containing 0.1 M ammonium sulfate in the stationary phase. 3. Continuous fluorometric monitoring of the changes in intrinsic peptide fluorescence. 4. Continuous fluorometric determination of NADH oxidation in a coupled enzyme assay.
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