Methods for analyzing the role of phospholipase A₂ enzymes in endosome membrane tubule formation.
Methods for analyzing the role of phospholipase A₂ enzymes in endosome membrane tubule formation.
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DOI：
10.1016/bs.mcb.2015.05.005
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发表时间：
2015
影响因子：
--
通讯作者：
Brown WJ
中科院分区：
文献类型：
--
作者：
Kalkofen DN;de Figueiredo P;Brown WJ
哺乳动物内体隔间的货物出口通常涉及膜小管，可溶性和膜结合的货物被分离到膜小管中，以便随后在细胞内运输。这些膜小管是高度动态的，它们的形成是由各种内小体相关蛋白介导的。然而，对这些膜小管如何在时间或空间上进行调节知之甚少，因此很可能会发现和分析其他与小管相关的蛋白。因此，对细胞生物学家来说，研究内吞体膜微管的生物发生和调控的方法将是很有价值的。在本章中，我们描述了使用无细胞、体外重建分析和活体图像分析工具来研究这一过程的方法。
Cargo export from mammalian endosomal compartments often involves membrane tubules, into which soluble and membrane-bound cargos are segregated for subsequent intracellular transport. These membrane tubules are highly dynamic and their formation is mediated by a variety of endosome-associated proteins. However, little is known about how these membrane tubules are temporally or spatially regulated, so other tubule-associated proteins are likely to be discovered and analyzed. Therefore, methods to examine the biogenesis and regulation of endosome membrane tubules will prove to be valuable for cell biologists. In this chapter, we describe methods for studying this process using both cell-free, in vitro reconstitution assays, and in vivo image analysis tools.