概括
研究人员分离了牛离子依赖的曼诺6酸盐 (CD Man-6-P) 受体cDNA克隆,通过表达一种特异性结合phosphomannan-Sepharose的蛋白质来确认它们的功能.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 曼6酸盐 (Man-6-P) 受体对于溶解体酶向至关重要.
- 存在两种不同的Man-6-P受体:阴阳系依赖 (CD) 和阴阳系独立 (CI).
- 了解CDMan-6-P受体的结构对于阐明其功能至关重要.
研究的目的:
- 为了分离和描述编码牛46kDaCDMan-6-P受体的cDNA克隆.
- 为了确认克隆受体的功能活性.
- 为了确定CD Man-6-P受体的结构特征.
主要方法:
- 隔离了编码牛CDMan-6-P受体的cDNA克隆.
- 在Xenopus laevis卵细胞中的受体mRNA的翻译.
- 使用索曼南-赛法洛斯的结合测试.
- 氨基酸序列分析.氨基酸序列分析.
- 微体膜的素消化.
- 识别N-链接的糖化位.
主要成果:
- 获得了牛CDMan-6-P受体的完整cDNA序列.
- 表达的蛋白质证明了与索曼南-酸盐的特定结合,证实了功能性.
- 推导的279个氨基酸序列揭示了具有信号序列和跨膜域的单个多.
- 受体是一种具有额外细胞质N端的跨膜蛋白.
- 外细胞质域与CI Man-6-P受体的重复域具有同质性.
结论:
- 孤立的cDNA克隆编码了一个功能性的牛CDMan-6-P受体.
- CD Man-6-P受体是一种具有明显的细胞外质域的跨膜蛋白.
- 结构上的相似性表明CD和CIMan-6-P受体之间保留了功能作用.
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