Electrophoretic purification of the alpha and beta subunits of phosphorylase kinase and evidence in support of the deduced amino acid sequences

✍ Scribed by Dr. John W. Crabb; William R. Harris; Charles M. Johnson; Theodore G. Sotiroudis; Carl C. Kuhn; Ludwig M. G. Heilmeyer Jr.

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 796 KB
• Volume: 11
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150110206

✦ Synopsis

Electrophoretic purification of the alpha and beta subunits of phosphorylase kinase and evidence in support of the deduced amino acid sequences

A simple, rapid sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method is presented for isolating the a, a' and p subunits of rabbit muscle phosphorylase kinase. The SDS-PAGE procedure can yield milligram amounts of a and fl from a single preparative gel and also allows isolation of the a' isozyme free of a.

Notably the method provides the purified subunits in a form amenable to structural analysis. Edman degradation of a and a' reveal identical NH,-terminal structures.

Amino acid analysis of the electrophoretically purified c1 and p subunits are in good agreement with their deduced primary structures. The amino acid sequence of 488 residues in a and 7 13 residues in p were determined by gas phase Edman degradation.

The data support the recently deduced primary structures of a (Zander et al., Proc.