High performance liquid chromatography purification and structural characterization of the subunits of rabbit muscle phosphorylase kinase

摘要: Abstract A rapid reverse-phase high performance liquid chromatography (HPLC) method is presented for isolating the alpha, beta, gamma, and delta subunits of rabbit muscle phosphorylase kinase. The HPLC separation allows micropreparative purification all with 66-88% recoveries. Relative molecular weights as determined by sodium dodecyl sulfate gel electrophoresis in 4, 5, 7, 10% acrylamide are alpha 132,000, alpha' 127,000, beta 113,000 gamma 43,000. Amino acid compositions reported purified subunits. contains about 2 mol endogenous phosphate/mol protein each contain 1 protein. Despite identity calmodulin, essentially no protein-bound phosphate was found associated bovine brain calmodulin. Holophosphorylase kinase 20 first NH2-terminal sequence analyses were Tarr manual Edman degradation. Within 23 residues ( TRDAALPGSHSTHGFYENYESKE . ) there six identities one conservative interchange catalytic subunit cAMP-dependent 17 MRSRSNSGVRLDSYARL exhibit transforming from Rous sarcoma virus (Schmidt- Rupin strain) provided a single gap inserted src gene product. Further structural information required to evaluate significance these similarities. has blocked NH2 terminus.