Biogenesis of phagolysosomes proceeds through a sequential series of interactions with the endocytic apparatus.
Biogenesis of phagolysosomes proceeds through a sequential series of interactions with the endocytic apparatus.
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DOI：
10.1083/jcb.124.5.677
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发表时间：
1994-03
期刊：
影响因子：
--
通讯作者：
Griffiths G
中科院分区：
文献类型：
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作者：
Desjardins M;Huber LA;Parton RG;Griffiths G
我们研究了在J-774巨噬细胞中吞噬体转化为吞噬溶酶体过程中发生的修饰。使用低密度乳胶珠作为吞噬体的标记物（乳胶珠隔室，LBC）允许通过在简单的蔗糖梯度上浮选来分离这些细胞器。二维凝胶电泳，免疫细胞化学和生化分析已被用来表征LBC的组成在不同的时间点后，他们的形成，以及他们的相互作用与细胞器的内吞途径。我们的研究结果表明，LBC获得和失去各种标志物转化为吞噬溶酶体。其中包括小GTP酶的rab家族成员以及lamp家族蛋白。LBC的灯2，与晚期内吞结构相关的膜蛋白的转移，被证明是微管依赖性的。视频显微镜显示，新形成的吞噬体参与快速多次接触的内吞途径的晚期组件。总的来说，这些观察结果表明，吞噬溶酶体的形成是一个高度动态的过程，涉及逐步和规范的收购标志物从内吞细胞器。
We have examined the modifications occurring during the transformation of phagosomes into phagolysosomes in J-774 macrophages. The use of low density latex beads as markers of phagosomes (latex bead compartments, LBC) allowed the isolation of these organelles by flotation on a simple sucrose gradient. Two-dimensional gel electrophoresis, immunocytochemistry, and biochemical assays have been used to characterize the composition of LBC at different time points after their formation, as well as their interactions with the organelles of the endocytic pathway. Our results show that LBC acquire and lose various markers during their transformation into phagolysosomes. Among these are members of the rab family of small GTPases as well as proteins of the lamp family. The transfer of the LBC of lamp 2, a membrane protein associated with late endocytic structures, was shown to be microtubule dependent. Video-microscopy showed that newly formed phagosomes were involved in rapid multiple contacts with late components of the endocytic pathway. Collectively, these observations suggest that phagolysosome formation is a highly dynamic process that involves the gradual and regulated acquisition of markers from endocytic organelles.