Isolation of functional, coated, endocytic vesicles
Isolation of functional, coated, endocytic vesicles
复制标题
功能性、包被、内吞性囊泡的分离
作者：
Philip G. Woodman;G. Warren
用[125I]转铁蛋白短暂内化来标记被包被的内吞噬囊泡，然后用2H2O和2H2O/Ficoll密度梯度联合纯化。通过测定内吞细胞器的融合来监测纯化，从而分离出功能性囊泡。分离的囊泡具有网格蛋白包被囊泡的所有特性，富含网格蛋白包被的主要成分，并且不被1 M Tris-HCl或未包被的atp酶包被。通过无细胞测定，近一半的标记囊泡能够参与随后的融合事件。融合具有特异性，需要能量和细胞质，对N-乙基马来酰亚胺敏感。
Brief internalization of [125I]transferrin was used to label coated endocytic vesicles, which were then purified using a combination of 2H2O and 2H2O/Ficoll density gradients. Purification was monitored using an assay measuring fusion of endocytic organelles, so as to isolate functional vesicles. Isolated vesicles had all the properties of clathrin-coated vesicles, being enriched for the major components of clathrin coats and uncoated by either 1 M Tris-HCl or an uncoating ATPase. Nearly half of the labeled vesicles were able to participate in subsequent fusion events, as measured by the cell-free assay. Fusion was specific, requiring energy and cytosol, and being sensitive to N- ethyl maleimide.
纯化囊泡运输所需的三种相关外周膜蛋白。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Clary,DO;Rothman,JE
通讯作者：
Rothman,JE