Purification of clathrin heavy and light chain from Dictyostelium discoideum.
Purification of clathrin heavy and light chain from Dictyostelium discoideum.
复制标题
从盘基网柄菌中纯化网格蛋白重链和轻链。
DOI：
10.1006/prep.1997.0793
复制
发表时间：
1997
期刊：
影响因子：
--
通讯作者：
O'Halloran,TJ
中科院分区：
文献类型：
--
作者：
Riddelle-Spencer,KS;O'Halloran,TJ
Clathrin, a protein important for endocytosis, is a hexamer composed of three heavy chains and three light chains. We report here the purification scheme used to isolate the clathrin protein from the simple eukaryote, Dictyostelium discoideum. Using a combination of differential centrifugation and column chromatography, we isolated approximately 2 mg of clathrin triskelions from 150-200 g of Dictyostelium cells. One additional step purified the 30-kDa clathrin light chain to homogeneity. Glycerol gradient centrifugation was used to determine an S value of 7.9 for purified clathrin. Rotary shadowed images of Dictyostelium clathrin revealed trimeric molecules with extended legs measuring 48+/-5 nm, similar in length to the legs of mammalian and yeast clathrin triskelions. The single clathrin light chain proved resistant to heat treatment, a property also similar to light chains from other species. The conservation of these physical properties in Dictyostelium clathrin demonstrates the potential of this model organism for the study of clathrin structure and function.
