[对虾组织中依赖环鸟苷酸的蛋白激酶的部分纯化及性质]

[Partial purification and properties of cGMP-dependent protein kinase from prawn tissue].

作者信息

Safarov N S, Musaev Z Sh, Baba-Zade S N, Sadykhov S T, Mekhtiev N Kh

出版信息

Biokhimiia. 1982 Jun;47(6):945-9.

Abstract

Using ion-exchange chromatography and gel filtration, cGMP-dependent protein kinase was purified from prawn tissues 220-fold with a yield of activity of 12%. The apparent Ka values for cGMP, cAMP and 8-Br-cGMP are 1 . 10(-7), 5 . 10(-6) and 5 . 10(-8) M, respectively; the apparent Km values for ATP in the presence of cGMP is 9 . 10(-6) M. The cGMP-stimulated protein kinase activity was observed only in the presence of SH-compounds and high Mg2+ concentrations (500-100 mM). The protein kinase demonstrated a broad pH optimum wih a maximum at pH 6.8-7.2. The elution volume of the enzyme during gel filtration corresponded to a globular protein with molecular weight of 140,000.

摘要

利用离子交换色谱法和凝胶过滤法，从对虾组织中纯化出依赖环鸟苷酸的蛋白激酶，纯化倍数为220倍，活性回收率为12%。环鸟苷酸（cGMP）、环腺苷酸（cAMP）和8-溴环鸟苷酸（8-Br-cGMP）的表观解离常数（Ka）值分别为1×10⁻⁷、5×10⁻⁶和5×10⁻⁸M；在存在cGMP的情况下，ATP的表观米氏常数（Km）值为9×10⁻⁶M。仅在存在巯基化合物和高浓度镁离子（500 - 100 mM）时，才观察到cGMP刺激的蛋白激酶活性。该蛋白激酶表现出较宽的pH最佳值，在pH 6.8 - 7.2时活性最高。凝胶过滤过程中该酶的洗脱体积对应于一种分子量为140,000的球状蛋白。