A cell-free biochemical complementation assay reveals complex and redundant cytosolic requirements for LRP endocytosis.

A cell-free biochemical complementation assay reveals complex and redundant cytosolic requirements for LRP endocytosis.
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无细胞生化互补测定揭示了 LRP 内吞作用复杂且冗余的胞质要求。

DOI:
10.1016/j.yexcr.2005.12.022
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发表时间:
2006
影响因子:
3.7
通讯作者:
Schmid,SandraL
Schmid,SandraL
中科院分区:
医学3区
文献类型:
--
作者:
Miwako,Ishido;Schmid,SandraL

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低密度脂蛋白受体相关蛋白(LRP)结合多种不同的配体,参与组成性内吞和信号转导。使用体外重建系统和一种新的生化互补试验,我们已经探讨了限制胞质的要求从分离的质膜LRP的内吞作用。我们发现,网格蛋白,AP 2和发动蛋白不支持有效的LRP摄取和牛脑细胞溶质(AS supt)的30%硫酸铵上清液馏分中存在的额外的因素是必需的。AS supt的分级显示,需要多个和冗余的因子来支持LRP内吞作用。其中Hsc 70、synaptojanin 1和CRMP-2经质谱鉴定。我们的数据表明,LRP,它承担几个不同的内吞基序在其胞质结构域,可能会使用多种途径的内吞作用在体外。
The low density lipoprotein receptor-related protein (LRP) binds multiple, distinct ligands and participates in constitutive endocytosis and signal transduction. Using an in vitro reconstitution system and a new biochemical complementation assay, we have explored the limiting cytosolic requirements for endocytosis of LRP from isolated plasma membranes. We find that clathrin, AP2 and dynamin do not support efficient LRP uptake and that additional factors present in a 30% ammonium sulfate supernatant fraction of bovine brain cytosol (AS supt) are required. Fractionation of the AS supt revealed that multiple and redundant factors are required to support LRP endocytosis. Among these, we identified Hsc70, synaptojanin1 and CRMP-2 by mass spectrometry. Our data suggest that LRP, which bears several distinct endocytic motifs in its cytoplasmic domain, may use multiple pathways for endocytosis in vitro.
DOI: 10.1038/nature01020
发表时间: 2002-09-26
期刊: NATURE
影响因子: 64.8
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形成网格蛋白和 COP 包被的运输囊泡的生化要求。
DOI: --
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