PURIFICATION AND PROPERTIES OF 100-KD PROTEINS FROM COATED VESICLES AND THEIR RECONSTITUTION WITH CLATHRIN
PURIFICATION AND PROPERTIES OF 100-KD PROTEINS FROM COATED VESICLES AND THEIR RECONSTITUTION WITH CLATHRIN
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DOI：
10.1002/j.1460-2075.1984.tb02075.x
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发表时间：
1984-01-01
期刊：
影响因子：
11.4
通讯作者：
ROBINSON, MS
中科院分区：
文献类型：
--
作者：
PEARSE, BMF;ROBINSON, MS
布洛克脑包被囊泡含有至少6个100-kd [千道尔顿]多肽家族，其具有促进网格蛋白组装的性质。这些蛋白质从Triton X-100提取的包被囊泡中纯化，通过凝胶过滤和羟基磷灰石和DE-52纤维素层析的组合。三种主要的100 kd物质与化学计量量的50 kd多肽形成复合物。在交联这些复合物时，主要产物似乎含有2个100 kd和2个50 kd的多肽。这些100-kd/50-kd复合物将与低浓度的网格蛋白结合，以得到主要为桶大小的相对均匀的包衣群体。其他三种100 kd的多肽缺乏50-kd蛋白，但在相同条件下与网格蛋白结合，产生宽范围大小的包衣，包括桶、截短的二十面体和直径> 100 nm的颗粒。当网格蛋白笼用饱和量的100-kd/50-kd复合物重新组装并通过EM研究时，额外的蛋白质似乎遵循网格蛋白多面体的基本几何形状，部分填充在笼结构的多边形面中。饱和度似乎需要. apprx。每个网格蛋白三聚体3个100-kd多肽分子。
Bullock brain coated vesicles contain a family of at least 6 100-kd [kilodalton] polypeptides which have the property of promoting clathrin assembly. These proteins were purified from Triton X-100-extracted coated vesicles by a combination of gel filtration and chromatography on hydroxylapatite and DE-52 cellulose. Three major 100-kd species occur as complexes with a stoichiometric amount of a 50-kd polypeptide. On cross-linking these complexes, the chief products appear to contain 2 polypeptides of 100 kd and 2 of 50 kd. These 100-kd/50-kd complexes will polymerize with low concentrations of clathrin to give a relatively homogeneous population of coats predominantly of the barrel size. Three other polypeptides of 100 kd lack the 50-kd protein but polymerize with clathrin under the same conditions to yield coats of a wide range of sizes including barrels, truncated icosahedra and particles of > 100 nm diameter. When clathrin cages are reassembled with a saturating amount of 100-kd/50-kd complexes and studied by EM, the additional proteins appear to follow the underlying geometry of the clathrin polyhedra, partially filling in the polygonal faces of the cage structures. Saturation appears to require .apprx. 3 molecules of 100-kd polypeptide per clathrin trimer.