B-50/GAP-43 potentiates cytoskeletal reorganization in raft domains
B-50/GAP-43 potentiates cytoskeletal reorganization in raft domains
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DOI：
10.1006/mcne.1999.0775
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发表时间：
1999-08-01
影响因子：
3.5
通讯作者：
Schotman, P
中科院分区：
文献类型：
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作者：
Aarts, LHJ;Verkade, P;Schotman, P
B-50 （GAP-43）是一种神经膜相关蛋白，与神经突生长和引导有关。用B-50 cDNA稳定转染鼠成纤维细胞后，我们观察到B-50免疫反应性在扁平的静息细胞中分散分布。相反，运动细胞在褶皱膜的前缘显示出高浓度的B-50，与肌动蛋白聚合一致。对瞬时转染B-50/EGFP的Rat1成纤维细胞的延时研究显示，大泡起源于褶膜。这些大囊泡（小泡细胞）中Thy-1（一种gpi锚定蛋白）呈阳性，但rab5（一种早期内体标志物）呈阴性。在初级海马神经元中，B-50也完全与筏标记物Thy-1共定位。抗体介导的海马神经元中Thy-1的交联导致细胞内蛋白B-50重新分布到Thy-1免疫阳性的膜斑块，而syntaxin主要被排除在这些斑块之外，表明B-50与筏相关。
B-50 (GAP-43) is a neural, membrane-associated protein that has been implicated in neurite outgrowth and guidance. Following stable transfection of Rat1 fibroblasts with B-50 cDNA we observed a dispersed distribution of B-50 immunoreactivity in flattened resting cells. In contrast, motile cells exhibited high concentrations of B-50 at the leading edge of ruffling membranes, coinciding with actin polymerization. Time-lapse studies on Rat1 fibroblasts transiently transfected with B-50/EGFP revealed that large vesicles originated from the ruffling membranes. These targe vesicles (pinocytes) were found positive for Thy-1, a GPI-anchored protein, but negative for rab-5, an early endosome marker. In primary hippocampal neurons B-50 also colocalized completely with the raft marker Thy-1. Antibody-mediated cross-linking of Thy-1 in hippocampal neurons resulted in a redistribution of the intracellular protein B-50 to Thy-1-immunopositive membrane patches, whereas syntaxin was mainly excluded from the patches, showing that B-50 is associated with rafts.