Andersen P H, Geisler A
Acta Pharmacol Toxicol (Copenh). 1983 Jul;53(1):49-56. doi: 10.1111/j.1600-0773.1983.tb01867.x.
A calmodulin-sensitive phosphodiesterase was purified from bovine brain. The purification procedure involved ammonium sulphate fractionation, two chromatographic steps on DEAE-cellulose, gel-filtration on Sephadex G-200, and finally one DEAE-cellulose run, and gave a 2300-fold purification. The purified phosphodiesterase had a Vmax for cyclic AMP of 126 mumol/mg protein X min. and was activated 8-fold by addition of calmodulin and calcium. According to SDS-electrophoresis the purified enzyme contained one major peptide of 59,000 daltons, but the preparation was not homogeneous. The enzyme was characterized kinetically and with regard to the effect of cations, pH temperature, and nucleotides. Furthermore, the influence in vitro on enzyme activity of several classes of drugs, e.g. antidepressants, neuroleptics, antiallergics, platelet inhibitors, and some "reference phosphodiesterase inhibitors" was investigated.
从牛脑中纯化出一种钙调蛋白敏感的磷酸二酯酶。纯化过程包括硫酸铵分级分离、在DEAE-纤维素上进行两步色谱分离、在Sephadex G-200上进行凝胶过滤，最后再进行一次DEAE-纤维素柱层析，纯化倍数达2300倍。纯化后的磷酸二酯酶对环磷酸腺苷（cAMP）的最大反应速度（Vmax）为126 μmol/mg蛋白质·分钟，添加钙调蛋白和钙后活性提高8倍。根据十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-电泳）结果，纯化后的酶含有一条主要的59,000道尔顿的肽链，但该制剂并不均一。对该酶进行了动力学特性以及阳离子、pH值、温度和核苷酸影响方面的研究。此外，还研究了几类药物，如抗抑郁药、抗精神病药、抗过敏药、血小板抑制剂以及一些“参考磷酸二酯酶抑制剂”在体外对酶活性的影响。
