Chun Y S, Shima H, Nagasaki K, Sugimura T, Nagao M
Carcinogenesis Division, National Cancer Center Research Institute, Tokyo, Japan.
Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3319-23. doi: 10.1073/pnas.91.8.3319.
Protein phosphatase 1 gamma 2 (PP1 gamma 2) is a testis-specific isotype of the protein-serine/threonine-phosphatase type 1 catalytic subunit. Three native forms of PP1 gamma 2 were detected in a crude fraction of rat testis by electrophoresis in a nondenaturing polyacrylamide gel. We purified a major native form of PP1 gamma 2 to homogeneity by successive column chromatography on Mono Q-Sepharose, EAH-agarose, protamine-agarose, and G3000SW and by electrophoresis in a nondenaturing polyacrylamide gel. The G3000SW-purified PP1 gamma 2 native form had an apparent molecular mass of 170 kDa. The purified holoenzyme from nondenaturing polyacrylamide gel was composed of the catalytic subunit and two noncatalytic subunits, of 78 kDa and 55 kDa. Partial amino acid sequence analysis of the 78-kDa protein suggested that it is the 78-kDa glucose-regulated protein, a member of the 70-kDa heat shock protein family. The 78-kDa protein may possibly function as a chaperone or by confining substrate specificity of PP1 gamma 2.
蛋白磷酸酶1γ2(PP1γ2)是蛋白丝氨酸/苏氨酸磷酸酶1催化亚基的睾丸特异性同种型。通过在非变性聚丙烯酰胺凝胶中进行电泳,在大鼠睾丸的粗提物中检测到三种天然形式的PP1γ2。我们通过在Mono Q-琼脂糖凝胶、EAH-琼脂糖、鱼精蛋白-琼脂糖和G3000SW上连续柱层析以及在非变性聚丙烯酰胺凝胶中进行电泳,将一种主要的天然形式的PP1γ2纯化至同质。经G3000SW纯化的PP1γ2天然形式的表观分子量为170 kDa。从非变性聚丙烯酰胺凝胶中纯化的全酶由催化亚基和两个非催化亚基组成,分子量分别为78 kDa和55 kDa。对78 kDa蛋白质的部分氨基酸序列分析表明,它是78 kDa葡萄糖调节蛋白,属于70 kDa热休克蛋白家族的一员。78 kDa蛋白质可能作为伴侣蛋白发挥作用,或者通过限制PP1γ2的底物特异性来发挥作用。