鸡延胡索酸酶。I. 纯化与特性鉴定。

Chicken fumarase. I. Purification and characterization.

作者信息

Reyns C, Léonis J, Schlusselberg J

出版信息

Biochimie. 1975;57(2):123-9. doi: 10.1016/s0300-9084(75)80161-1.

Abstract

Fumarase from chicken heart is purified 400 times from the crude muscle extract. The isolation procedure includes ammonium sulfate fractionations, Bio-Gel P-300 column chromatography and electrofocusings on pH-gradients from pH 3 to 10 and from pH 7 to 9. Chicken fumarase behaves as an homogeneous protein in sedimentation, diffusion and electrofocusing studies; the protein possesses a single amino-terminal residue: lysine. The analysis of the CD and ORD spectra suggests the presence of 60-65 p. cent of alpha-helix, 0 - 5 p. cent of beta-structure with the remaining portions of the protein in an unordered conformation. Chicken fumarase is found to be composed of 4 subunits of identical molecular weight (51.000) and devoid of disulfide bridges. Finally, the physicochemical properties of chicken fumarase are compared with those of the porcine enzyme.

摘要

鸡心延胡索酸酶从粗肌肉提取物中纯化了400倍。分离过程包括硫酸铵分级分离、Bio-Gel P-300柱层析以及在pH 3至10和pH 7至9的pH梯度上进行等电聚焦。在沉降、扩散和等电聚焦研究中，鸡延胡索酸酶表现为一种均一的蛋白质；该蛋白质具有单个氨基末端残基：赖氨酸。圆二色光谱（CD）和旋光色散光谱（ORD）分析表明，α-螺旋占60 - 65%，β-结构占0 - 5%，蛋白质的其余部分呈无序构象。发现鸡延胡索酸酶由4个分子量相同（51,000）的亚基组成，且不含二硫键。最后，将鸡延胡索酸酶的物理化学性质与猪酶的性质进行了比较。