Evidence that a cAMP binding protein from Dictyostelium discoideum carries S-adenosyl-L-homocysteine hydrolase activity.
A cAMP-adenosine binding protein partially purified from exponentially growing Dictyostelium discoideum cells carries S-adenosyl-L-homocysteine (SAH) hydrolase activity. This protein is present throughout the developmental cycle and has many properties in common with a cAMP binding activity previously reported from this laboratory (Gunzburg and Véron, 1981). Direct binding measurements with radioactive ligands indicate a dissociation constant of 0.2 microM for adenosine and 9 nM for cAMP, a value in good agreement with measurements of the rate constants for cAMP binding (k+1 = 2.4 X 10(4) M-1 sec-1) and dissociation (k-1 = 1.1 X 10(-4) sec-1). The binding of cAMP is completely abolished in the presence of 1 microM adenosine; a maximum 60 per cent inhibition of adenosine binding can be achieved with cAMP concentrations as high as 0.1 microM, suggesting that at least some of the cAMP and adenosine binding sites are not identical. The protein has a sedimentation coefficient of 9.2S and a native molecular weight of 190,000, as judged by gel filtration. Labeling with the photoaffinity ligand 8-azido-[3H]-cAMP followed by SDS polyacrylamide gel electrophoresis results in a single band of 47,000 MW, suggesting that the protein may be a tetramer. The physiological importance of the protein and its association with SAH hydrolase activity is discussed in relation to a possible role in the regulation of protein and phospholipid methylation that occurs during chemotaxis.
D000241:Adenosine / Q000378:metabolism; D043383:Adenosylhomocysteinase; D002352:Carrier Proteins / Q000032:analysis*; D002852:Chromatography, Ion Exchange; D000242:Cyclic AMP / Q000378:metabolism; D002373:Cyclic AMP Receptor Protein; D004023:Dictyostelium / Q000032:analysis*; D006867:Hydrolases / Q000378:metabolism*; D008970:Molecular Weight
D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D000242:Cyclic AMP; D006867:Hydrolases; D043383:Adenosylhomocysteinase; D000241:Adenosine