>Several steps of purification (octyl-Sepharose chromatography,Blue Sepharose 6B Chromatography and sucrose density gradient centrifugation) led to a highly purified aggregate of the enzymes, 3′, 5′-cyclic-nucleotide phosphodiesterase (PDE) and nucleotidase. The purfied enzyme aggregate showed an S value of 7.3 (SE±0.3, n = 10). Further analysis by SDS-polyacrylamide gel electrophoresis (PAGE) reavealed two proteins near 67 and 60 kDa. Dissocation of the 7.3 S enzyme aggregate showed a 3.6 S PDE form and a nucleotidase form at 4.2 S. Additionally, higher S value forms of the necleotidase up to 17 S have been observed. Apparently, they had formed by self-association. SDS-PAGE of the 17 S nucleotidase form showed only one hand at 67 kDa. This was taken as evidence for the homogenity of the 17 S nucleotidase form and the self-association of the nuleotidase after dissociation from the 7.3 S enzyme aggregate. Furthermore, from this it could be concluded that the 67 kDa protin of the 7.3 S enzyme aggregate should be identified with the nucleotidase, and thus the 60 kDa band represents the PDE.
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机译：几个纯化步骤（辛基-琼脂糖层析，蓝琼脂糖6B色谱和蔗糖密度梯度离心法）产生了高度纯化的酶，3'，5'-环核苷酸磷酸二酯酶（PDE）和核苷酸酶的聚集体。纯化的酶聚集体的 S 值为7.3（SE±0.3， n = 10）。通过SDS-聚丙烯酰胺凝胶电泳（PAGE）进行的进一步分析去除了67和60 kDa附近的两种蛋白质。 7.3 S酶聚集体的解离显示3.6 S PDE形式和4.2 S处的核苷酸酶形式。此外，还观察到了最高17 S的核苷酸酶的 S 值形式。显然，他们是通过自我交往形成的。 17 S核苷酸酶形式的SDS-PAGE在67 kDa时只显示了一只手。这被作为17 S核苷酸酶形式的同质性和从7.3 S酶聚集体解离后核酸酶的自缔合的证据。此外，由此可以得出结论，应该用核苷酸酶鉴定7.3 S酶聚集体的67 kDa蛋白质，因此60 kDa带代表PDE。
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