Walter U, Miller P, Wilson F, Menkes D, Greengard P
J Biol Chem. 1980 Apr 25;255(8):3757-62.
A guanosine 3':5'-monophosphate (cGMP)-dependent protein kinase was purified from bovine lung using 8-(6-aminohexylamino)-cAMP-Sepharose. The activity of the purified enzyme was highly dependent on cGMP using histone f2b as a substrate. The self-phosphorylation of the purified enzyme was strongly inhibited by cGMP and not significantly affected by cAMP. A precipitating antiserum prepared in rabbits against the cGMP-dependent protein kinase specifically inhibited the histone kinase activity and the self-phosphorylation of the purified cGMP-dependent protein kinase without affecting the cGMP binding site. This antiserum also specifically inhibited the phosphorylation of the endogenous substrate proteins by endogenous cGMP-dependent protein kinase in smooth muscle membranes, but did not cross-react detectably with catalytic subunit or regulatory subunit of type I or type II cAMP-dependent protein kinase. Conversely, anti-sera against the regulatory subunit of type I or type II cAMP-dependent protein kinase did not cross-react detectably with cGMP-dependent protein kinase. The substantial differences between the immunological properties of the cGMP-dependent and cAMP-dependent protein kinases suggest that these two enzymes have distinct physiological roles.
使用8-(6-氨基己基氨基)-cAMP-琼脂糖从牛肺中纯化出一种3':5'-环磷酸鸟苷（cGMP）依赖性蛋白激酶。以组蛋白f2b为底物时，纯化酶的活性高度依赖于cGMP。纯化酶的自身磷酸化受到cGMP的强烈抑制，而cAMP对其无显著影响。用兔制备的针对cGMP依赖性蛋白激酶的沉淀抗血清可特异性抑制纯化的cGMP依赖性蛋白激酶的组蛋白激酶活性和自身磷酸化，且不影响cGMP结合位点。该抗血清还可特异性抑制平滑肌膜中内源性cGMP依赖性蛋白激酶对内源底物蛋白的磷酸化，但与I型或II型cAMP依赖性蛋白激酶的催化亚基或调节亚基无明显交叉反应。相反，针对I型或II型cAMP依赖性蛋白激酶调节亚基的抗血清与cGMP依赖性蛋白激酶无明显交叉反应。cGMP依赖性蛋白激酶和cAMP依赖性蛋白激酶免疫特性的显著差异表明这两种酶具有不同的生理作用。
