Intracellular glycosylphosphatidylinositols accumulate on endosomes: toxicity of alpha-toxin to Leishmania major.
Intracellular glycosylphosphatidylinositols accumulate on endosomes: toxicity of alpha-toxin to Leishmania major.
复制标题
细胞内糖基磷脂酰肌醇在内体上积聚：α-毒素对利什曼原虫的毒性。
DOI：
10.1128/ec.4.3.556-566.2005
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Mensa-Wilmot,Kojo
中科院分区：
文献类型：
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作者：
Zheng,Zhifeng;Tweten,RodneyK;Mensa-Wilmot,Kojo
糖基磷脂酰肌醇（GPIs）是真核生物中普遍存在的糖脂。在原生动物硕大利什曼原虫中，GPIs“游离”或共价连接到蛋白质上（例如，gp 63）和多糖。虽然在质膜上检测到一些游离的GPIs，但GPIs在细胞内积累的特定位点在大多数细胞中是未知的，尽管糖脂在分泌系统内合成。在此，我们描述了一个协议，用于确定GPI积累的细胞内网站使用α-毒素（从梭菌败血症）。α-毒素与gp 63和来自mL的GPIs结合。少校在去除GPI锚定的大分子（例如，gp 63）从固定细胞的质膜上分离。内体是GPI在L.少校在内质网中检测到GPI-少的gp 63。在对活寄生虫的研究中，α毒素杀死了L。major的50%致死浓度为0.77 nM。
Glycosylphosphatidylinositols (GPIs) are ubiquitous glycolipids in eukaryotes. In the protozoanLeishmania major, GPIs occur “free” or covalently linked to proteins (e.g., gp63) and polysaccharides. While some free GPIs are detected on the plasma membrane, specific sites where GPIs accumulate intracellularly are unknown in most cells, although the glycolipids are synthesized within the secretory system. Herein, we describe a protocol for identifying intracellular sites of GPI accumulation by using alpha-toxin (fromClostridium septicum). Alpha-toxin bound to gp63 and GPIs fromL. major. Intracellular binding sites for alpha-toxin were determined in immunofluorescence assays after removal of GPI-anchored macromolecules (e.g., gp63) from the plasma membrane of fixed cells by using detergent. Endosomes were a major site for GPI accretion inL. major. GPI-less gp63 was detected at the endoplasmic reticulum. In studies with live parasites, alpha-toxin killedL. majorwith a 50% lethal concentration of 0.77 nM.