A comparative study of histone deacetylases of plant, fungal and vertebrate cells
A comparative study of histone deacetylases of plant, fungal and vertebrate cells
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DOI：
10.1016/0167-4838(96)00069-6
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发表时间：
1996-09-05
期刊：
影响因子：
--
通讯作者：
Loidl, P
中科院分区：
文献类型：
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作者：
Lechner, T;Lusser, A;Loidl, P
可逆核心组蛋白乙酰化的酶平衡由两种酶活性维持，即组蛋白乙酰转移酶和组蛋白脱乙酰酶（HD）。这些酶活性以多种酶形式存在。本报告描述了方法，以提取不同的HD形式从三种生物，发芽玉米胚，粘菌绒泡菌多头绒泡菌，和鸡红细胞，它提供了数据的色谱分离和部分纯化的HD形式。在萌发的玉米胚中，可以区分三种HD（HD 1-A、HD 1-B、HD 2）;根据它们对pH、温度和各种离子的依赖性以及动力学参数（核心组蛋白的k（M））和各种化合物的抑制来表征HD 1-A、HD 1-B和HD 2。相同的参数进行了研究，为相应的酶的多头绒泡菌，成熟和未成熟的鸡红细胞。基于这些结果，建立了不同酶形式的最佳测定条件。动力学数据显示，玉米组蛋白脱乙酰酶HD 1-B峰经Q-Sepharose层析部分纯化后为异质性峰，由两个组蛋白结合位点组成，这两个位点对纯化的核心组蛋白的亲和力差异显著。在聚赖氨酸琼脂糖上的优化亲和层析确实表明，前定义的脱乙酰酶HD 1-B可以清楚地分离成两个单独的HD酶形式。组蛋白去乙酰化酶的高度多样性强调了这些酶对于核心组蛋白乙酰化的复杂调节的重要性。
The enzymatic equilibrium of reversible core histone acetylation is maintained by two enzyme activities, histone acetyltransferase and histone deacetylase (HD). These enzyme activities exist as multiple enzyme forms. The present report describes methods to extract different HD-forms from three organisms, germinating maize embryos, the myxomycete Physarum polycephalum, and chicken red blood cells; it provides data on the chromatographic separation and partial purification of HD-forms. In germinating maize embryos three HDs (HD1-A, HD1-B, HD2) can be discriminated; HD1-A, HD1-B, and HD2 were characterized in terms of their dependence on pH, temperature and various ions, as well as kinetic parameters (k(M) for core histones) and inhibition by various compounds. The same parameters were investigated for the corresponding enzymes of Physarum polycephalum, and mature and immature chicken erythrocytes. Based on these results, optimum assay conditions were established for the different enzyme forms. The kinetic data revealed that the maize histone deacetylase HD1-B peak after partial purification by Q-Sepharose chromatography was heterogeneous and consisted of two histone binding sites that differed significantly in their affinity for purified core histones. Optimized affinity chromatography on poly-Lysine Agarose indeed showed that the former defined deacetylase HD1-B can be separated clearly into two individual HD enzyme forms. The high multiplicity of histone deacetylases underlines the importance of these enzymes for the complex regulation of core histone acetylation.