An unusual adenosine cyclic 3',5'-phosphate-dependent protein kinase from Dictyostelium discoideum

Abstract

The CAMP-dependent protein kinase from Dictyostelium discoideum was extracted from cells at the stage of culmination. Less than 50% of the enzyme remains as a CAMP-dependent holoenzyme in the extracts, and the rest is recovered in the form of dissociated regulatory and catalytic subunits that were purified. The regulatory subunit is a monomeric protein of M, 42 000 that carries only one cAMP binding site (Kd = 3 nM). The catalytic subunit is also a monomer of M, 40000 with a sedimentation coefficient of 3.3S. The CAMP-dependent holoenzyme is a dimer consisting of one regulatory and one catalytic subunit, and the same structure is found for the holoenzyme reconstituted from the isolated subunits. Whereas cAMP binding to the regulatory subunit is independent of pH, both the catalytic activity and its ability to be inhibited by addition of regulatory subunit are increased very strongly between pH 5.5 and 7. The differences in molecular and catalytic properties of this CAMP-dependent protein kinase with those from mammalian origin are discussed in relation with the possibility that the enzyme from Dictyostelium represents an early form of the molecule in the evolutionary process.