title: Novel protein kinase, AUT-PK 85, isolated from adrenocortical carcinoma: purification and characterization.
abstract: We describe the purification to apparent homogeneity of a protein kinase (designated AUT-PK 85) from adrenocortical carcinoma 494, as evidenced by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The enzyme binds cyclic AMP (cAMP) and autophosphorylates but does not use histone, casein, or polysomes as substrates in the presence or absence of cAMP. Stoichiometry of phosphate incorporation was 0.71 mol/mol of enzyme. The enzyme was found to have a molecular weight of 85,000 based on gel filtration. The protein was composed of polypeptides having the same molecular weight 42,000, and thus it appears to consist of two subunits of equal size. The enzyme bound two cAMP molecules, indicating that each subunit binds one molecule of cAMP. The homogeneous enzyme did not inhibit the protein kinase activity of the free catalytic subunit of normal adrenal cAMP-dependent protein kinase under conditions such that recombination with the free regulatory subunit occurred. cAMP bound specifically to the enzyme with an apparent dissociation constant (cfKd) of 1.2 X 10(-8) M. Scatchard plot data indicated one type of binding sites for cAMP. The enzyme did not bind adenosine. This novel autophosphorylating, cAMP-binding, protein kinase may be a characteristic of certain adrenal neoplasms.
mesh_terms: D000306:Adrenal Cortex Neoplasms / Q000201:enzymology*; D000345:Affinity Labels; D000818:Animals; D002277:Carcinoma / Q000201:enzymology*; D002845:Chromatography / Q000379:methods; D000242:Cyclic AMP / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D046911:Macromolecular Substances; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000201:enzymology; D010766:Phosphorylation; D011485:Protein Binding; D011494:Protein Kinases / Q000302:isolation & purification*
chemical_list: D000345:Affinity Labels; D006657:Histones; D046911:Macromolecular Substances; D000242:Cyclic AMP; D011494:Protein Kinases