Belikoff E, Wong L J, Alberts B M
J Biol Chem. 1980 Dec 10;255(23):11448-53.
A concentrated, DNA-free extract of histone acetylase A was prepared from calf thymus tissues in two simple steps, which exploit the ability of polyethylene glycol to precipitate both nucleic acids and proteins from solutions containing high concentrations of salt (Alberts, B., and Herrick, G. (1971) Methods Enzymol. 21, 198-217). This extract was then chromatographed on four successive columns. The use of 75 microgram/ml of insulin as a carrier protein in all of these later steps, plus the inclusion of 1 M urea in some column buffers, has been useful in improving both the yield and reproducibility of the purification. The highly active enzyme obtained has a molecular weight of about 70,000, and the best fractions could be about 30% pure. Our data indicate that the acetylase A is only a very minor protein in cells, being present in perhaps a few thousand molecules per cell.
通过两个简单步骤从小牛胸腺组织中制备了浓缩的、不含DNA的组蛋白乙酰化酶A提取物，这利用了聚乙二醇从含有高浓度盐的溶液中沉淀核酸和蛋白质的能力（阿尔伯茨，B.，和赫里克，G.（1971年）《酶学方法》21，198 - 217）。然后将该提取物在四个连续的柱上进行色谱分析。在所有这些后续步骤中使用75微克/毫升的胰岛素作为载体蛋白，以及在一些柱缓冲液中加入1 M尿素，有助于提高纯化的产量和重现性。获得的高活性酶的分子量约为70,000，最佳级分的纯度可能约为30%。我们的数据表明，乙酰化酶A在细胞中只是一种非常次要的蛋白质，每个细胞中可能仅存在几千个分子。
