This chapter discusses the removal of phosphate from proteins by the reverse reaction. Covalently bound phosphate can be removed from proteins by simple chemical means or by enzymatically catalyzed reactions. Chemical means include mild alkali treatment to remove phosphate from serine and threonine residues through β-elimination reactions. However, such reactions are only one of the many effects of treating proteins with mild alkali. The lack of specificity of such methods makes them unsuitable for studying the effect of removing phosphate from specific sites in proteins, in particular the sites that are not alkali-labile. However, these methods have the advantage that they allow the possibility of activity measurement after a relatively mild and specific treatment to remove phosphate. The particular methods and conditions described in this chapter are designed for work with the cyclic nucleotide-dependent protein kinases. The principles involved are also applicable to many other purified protein kinases, which might effectively remove phosphate from proteins under appropriately designed conditions.
ASJC Scopus subject areas
- Molecular Biology