糖基化猪催乳素(G-PRL)的分离与性质研究。

Bollettino chimico farmaceutico Pub Date : 2004-05-01
F Ryszka, B Dolinska
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引用次数: 0

摘要

糖基化猪催乳素(G-PRL)的分离及性质研究——用70%的酸性丙酮从冻干的猪垂体后叶中提取粗催乳素,然后将提取液中的丙酮浓度提高到92%,进行蛋白质沉淀。将收到的沉淀物溶解于0.1 M醋酸中,催乳素在pH = 4.97的等电点沉淀。将催乳素沉淀溶解于pH = 7.50的磷酸盐缓冲液中,吸附在DEAE-Sephadex色谱柱上,在NaCl浓度为0.00-0.40 m范围内,用线性梯度洗脱,在NaCl浓度为0.19-0.25 m范围内洗脱分子量为24.0 kD的催乳素,在Sephadex G-100上采用凝胶过滤法将糖基化催乳素分为分子量为-55.8和17.4 kD的2个组分。24.0 kD催乳素及其糖基化形式均具有较高的生物活性。糖基化催乳素中含有甘露糖和焦糖。
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Isolation and properties of glycosylated pig prolactin (G-PRL).

Isolation and properties of glycosylated pig prolactin (G-PRL)--Crude prolacting was received via extraction from lyophilized hypophyses with acidic 70% acetone, and then by protein precipitation via increasing acetone concentration in the extract to 92%. Received precipitate was dissolved in 0.1 M acetic acid and prolactin was precipitated in the isoelectric point at the pH = 4.97. Prolactin precipitate was dissolved in phosphate buffer pH = 7.50 and adsorbed on the DEAE-Sephadex column, and then eluated with linear gradient at NaCl concentration range 0.00-0.40 M. Prolactin with molecular mass 24.0 kD was eluated at NaCl concentration 0.19-0.25 M. Using gel filtration method on Sephadex G-100 glycosylated prolactin was divided into 2 fractions with molecular mass equal--55.8 and 17.4 kD. Both 24.0 kD--prolactin and its glycosylated form were characterized by high biological activity. Glycosylated prolactin contains among others mannose and fucose.

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