Tanaka T, Kotani T, Ohtaki S, Nagai K, Tsuruta K, Mori N
Nihon Sanka Fujinka Gakkai Zasshi. 1987 Apr;39(4):579-85.
A NAD+-dependent 15-hydroxyprostaglandin dehydrogenase was purified from human term placenta using monoclonal antibody-coupled Sepharose affinity chromatography. The monoclonal antibodies were prepared against the molecule which was purified according to the method of Jarabak et al. About 4,800-fold purification was achieved with a 3% yield. The highest specific activity was 1,890 mU/mg. The molecular weight of the enzyme determined by gel filtration was 49,000. Polyacrylamide gel electrophoresis of the purified enzyme revealed the presence of two protein bands which were thought to be derived from the same enzyme molecule. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed two main bands of which the molecular weights were 28,000 to 30,000 and 52,000 to 56,000. Electrophoretic analyses suggest a labile oligomeric structure that may relate to the instability of the enzyme activity, which contradicts the monomer theory reported by Jarabak et al.
利用单克隆抗体偶联琼脂糖亲和色谱法从人足月胎盘纯化出一种NAD⁺依赖性15-羟基前列腺素脱氢酶。针对按照Jarabak等人的方法纯化得到的分子制备了单克隆抗体。纯化倍数达到约4800倍，产率为3%。最高比活性为1890 mU/mg。通过凝胶过滤测定的该酶分子量为49,000。纯化酶的聚丙烯酰胺凝胶电泳显示存在两条蛋白带，认为它们源自同一酶分子。十二烷基硫酸钠聚丙烯酰胺凝胶电泳显示两条主要条带，其分子量分别为28,000至30,000以及52,000至56,000。电泳分析表明存在一种不稳定的寡聚结构，这可能与酶活性的不稳定性有关，这与Jarabak等人报道的单体理论相矛盾。
