Identification of a proacrosin precursor in the cell-free translation of boar testicular poly(A)(+)-mRNA.
Identification of a proacrosin precursor in the cell-free translation of boar testicular poly(A)(+)-mRNA.
复制标题
公猪睾丸 Poly(A)( )-mRNA 无细胞翻译中顶体素前体的鉴定。
DOI：
10.1095/biolreprod44.2.332
复制
发表时间：
1991
影响因子：
3.6
通讯作者：
Polakoski,KL
中科院分区：
文献类型：
--
作者：
Yi,LS;Erbs,PA;Willand,JL;Polakoski,KL
采用无细胞翻译系统测定了猪原顶蛋白前体蛋白组分的分子质量。Poly(A)+-mRNA从新鲜切除的猪睾丸中提取到苯酚/氯仿中，在冷冻（-20°C）乙醇中沉淀，然后在无细胞的网状细胞裂解体系中翻译，并带有tran35s标签。通过SDS-PAGE和放射自显影对产物进行分析，发现了翻译蛋白的多个条带。Western blotting和特异的猪proacrosin多克隆抗体免疫沉淀均得到一个相对分子量约为64 000的主条带。这些结果表明，含有蛋白质和碳水化合物的原acrosin （Mr= 53 000 - 55 000）是由原acrosin前体分子的细胞加工产生的。
A cell-free translation system was used to determine the molecular mass of the protein component of precursor(s) to boar proacrosin. Poly(A)+-mRNA was extracted from freshly excised boar testis into phenol/chloroform, precipitated in chilled (-20°C) ethanol, then translated in a cell-free, reticulocyte lysate system with Tran35S-label. Analysis of the resulting products by SDS-PAGE followed by autoradiography demonstrated multiple bands of translated proteins. Both Western blotting and immunoprecipitation with a specific polyclonal antibody to boar proacrosin yielded a single major band with a relative molecular weight of approximately 64 000. These results suggest that proacrosin (Mr= 53 000–55 000), which contains both protein and carbohydrate moieties, results from the cellular processing of a proacrosin precursor molecule.