Morozov V E, Eronina T B, Andreeva I E, Silonova G V, Solov'eva N V
Biokhimiia. 1989 Mar;54(3):448-55.
Using DEAE-Toyopearl column chromatography, a preparation of pigeon skeletal muscle phosphorylase kinase was obtained in a state approaching homogeneity. The molecular mass of the native enzyme (1320 kDa) and the subunit formula (alpha beta gamma delta)4 are similar to those of rabbit and chicken counterparts. Both red and white pigeon skeletal muscle isozymes contain the alpha'-subunit instead of alpha. Gradient SDS-PAGE electrophoresis revealed small but well-reproducible differences in the molecular masses of rabbit, chicken and pigeon muscle beta- and gamma-subunits. The activity ratio at pH 6.8/8.2 is 0.06-0.15 for different preparations of phosphorylase kinase b. The activity of pigeon muscle phosphorylase kinase b is Ca2+-dependent. The [Ca2+]0.5 value at pH 7.0 is 20 microM, which exceeds that for the chicken muscle enzyme by two orders of magnitude. In the presence of Ca2+, pigeon phosphorylase kinase b is activated 4-fold by saturating concentrations of calmodulin and troponin C. Pigeon muscle phosphorylase b is activated 3-5-fold during autophosphorylation or phosphorylation by the catalytic subunit of cAMP-dependent protein kinase.
使用二乙氨基乙基琼脂糖凝胶柱色谱法，获得了接近均一状态的鸽骨骼肌磷酸化酶激酶制剂。天然酶的分子量（1320 kDa）和亚基组成（αβγδ）4与兔和鸡的相应酶相似。红肌和白肌鸽骨骼肌同工酶均含有α'-亚基而非α亚基。梯度SDS-PAGE电泳显示，兔、鸡和鸽肌肉β-和γ-亚基的分子量存在微小但可重复的差异。不同磷酸化酶激酶b制剂在pH 6.8/8.2时的活性比为0.06 - 0.15。鸽肌肉磷酸化酶激酶b的活性依赖于Ca2+。在pH 7.0时，[Ca2+]0.5值为20 μM，比鸡肌肉酶高出两个数量级。在Ca2+存在的情况下，饱和浓度的钙调蛋白和肌钙蛋白C可使鸽磷酸化酶激酶b的活性提高4倍。鸽肌肉磷酸化酶b在自磷酸化或被cAMP依赖性蛋白激酶催化亚基磷酸化过程中活性提高3 - 5倍。
