[25] Radiolabeling and detection methods for studying metabolism of regulatory subunit of cAMP-dependent protein kinase I in intact cultured cells

摘要: Publisher Summary This chapter discusses procedures for combining metabolic labeling with two-dimensional gel analysis to study the regulation of R 1 metabolism. Alterations in expression cAMP-dependent protein kinase subunits accompanying activation, tissue differentiation, cellular transformation, cell cycle traverse, or somatic mutations have generated interest subunit metabolism and its intracellular regulation. The most direct approach studying specific intact cells involves incorporation radiolabeled amino acids subsequent quantitation radioactivity associated interest. Protein modifications can be followed using radioisotopes modifying groups or, some modifications, by alterations electrophoretic mobility protein. There has been considerable success high-resolution polyacrylamide electrophoresis crude extracts resolve type I regulatory (R ) studies synthesis degradation cultured S49 mouse lymphoma cells.