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首页> 外文期刊>Life sciences >Lagenin, a novel ribosome-inactivating protein with ribonucleolytic activity from bottle gourd (Lagenaria siceraria) seeds.
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Lagenin, a novel ribosome-inactivating protein with ribonucleolytic activity from bottle gourd (Lagenaria siceraria) seeds.

机译：Lagenin，一种新型的核糖体失活蛋白，来自葫芦（Lagenaria siceraria）种子，具有核糖核酸分解活性。

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The seeds of Lagenaria siceraria (Family Cucurbitaceae) were extracted with water and the extract was lyophilized. The lyophilized extract was chromatographed on a DEAE-cellulose column in 10 mM Tris-HCl buffer (pH 7.2). The unadsorbed fraction was applied to an Affi-gel Blue gel column previously equilibrated with the same buffer. After removal of unadsorbed materials, the adsorbed proteins were eluted with 1.5 M NaCl in the Tris-HCl buffer. After dialysis the adsorbed fraction was loaded on a CM-Sepharose CL-6B column which had been equilibrated with and was eluted with the same buffer. After elution of unadsorbed proteins, the column was eluted with a gradient of 0-1 M NaCl in 10 mM Tris-HCl buffer (pH 7.2). The fraction eluting at about 0.55 M NaCl, which represented pure ribosome inactivating protein (RIP), inhibited cell-free translation in a rabbit reticulocyte system with an IC50 of 0.21 nM and exerted ribonuclease activity on yeast tRNA with an activity of 45 U/mg. The RIP was designated lagenin. It possessed a molecular weight of 20 kDa, smaller than the range of 26-32 kDa reported for other RIPs. The N-terminal sequence of lagenin exhibited a lesser extent of similarity to those of other Cucurbitaceae RIPs, characterized by a deletion of the first three amino acid residues and a replacement of the 4th (Phe), 17th (Phe), 18th (Ile) and 22nd (Arg) residues which are invariant in other RIPs.
机译：用水提取西洋参（Lagenaria siceraria）（葫芦科）的种子，并将提取物冻干。将冻干的提取物在DEAE-纤维素柱上在10mM Tris-HCl缓冲液（pH 7.2）中进行色谱分离。将未吸附的级分加到事先用相同缓冲液平衡过的Affi-gel Blue凝胶柱上。除去未吸附的物质后，将吸附的蛋白质在Tris-HCl缓冲液中用1.5 M NaCl洗脱。透析后，将吸附的级分加载到已经用相同缓冲液平衡并洗脱的CM-Sepharose CL-6B柱上。洗脱未吸附的蛋白质后，将色谱柱在10 mM Tris-HCl缓冲液（pH 7.2）中用0-1 M NaCl梯度洗脱。在约0.55 M NaCl上洗脱的级分代表纯核糖体失活蛋白（RIP），抑制兔网织红细胞系统中的无细胞翻译，IC50为0.21 nM，并在酵母tRNA上发挥核糖核酸酶活性，活性为45 U / mg 。 RIP被指定为lagenin。它的分子量为20 kDa，小于其他RIP报道的26-32 kDa范围。皂苷元的N-末端序列与其他葫芦科RIP的相似性较小，其特征是前三个氨基酸残基缺失，第4个（Phe），第17个（Phe），第18个（Ile）取代和在其他RIP中不变的第22个（Arg）残基。

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