Colocalization and nonrandom distribution of Kv1.3 potassium channels and CD3 molecules in the plasma membrane of human T lymphocytes
Colocalization and nonrandom distribution of Kv1.3 potassium channels and CD3 molecules in the plasma membrane of human T lymphocytes
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DOI：
10.1073/pnas.0438057100
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发表时间：
2003-03-04
影响因子：
11.1
通讯作者：
Damjanovich, S
中科院分区：
文献类型：
--
作者：
Panyi, G;Bagdány, M;Damjanovich, S
利用电子显微镜、激光共聚焦扫描显微镜和荧光共振能量转移技术研究了Kv1.3钾通道和CD 3分子的分布和横向组织。免疫金标记和电子显微镜显示，FLAG表位标记的Kv1.3通道（Kv1.3/FLAG）的分布显着不同的随机泊松分布在人T淋巴瘤细胞的质膜。共聚焦激光扫描显微镜图像显示，Kv1.3/FLAG通道和CD 3分子聚集在很大程度上重叠的膜区域。空间强度分布的互相关的数值分析得到了很高的相关系数（C = 0.64）。Kv1.3/FLAG和CD 3蛋白之间的分子接近度的不同层次水平被报道为高荧光共振能量转移效率（E = 51%）。这些发现暗示离子通道活性、膜电位和受体复合物功能的相互调节可能有助于免疫突触的正常功能。
Distribution and lateral organization of Kv1.3 potassium channels and CD3 molecules were studied by using electron microscopy, confocal laser scanning microscopy, and fluorescence resonance energy transfer. Immunogold labeling and electron microscopy showed that the distribution of FLAG epitope-tagged Kv1.3 channels (Kv1.3/FLAG) significantly differs from the stochastic Poisson distribution in the plasma membrane of human T lymphoma cells. Confocal laser scanning microscopy images showed that Kv1.3/FLAG channels and CD3 molecules accumulated in largely overlapping membrane areas. The numerical analysis of crosscorrelation of the spatial intensity distributions yielded a high correlation coefficient (C = 0.64). A different hierarchical level of molecular proximity between Kv1.3/FLAG and CD3 proteins was reported by a high fluorescence resonance energy transfer efficiency (E = 51%). These findings implicate that reciprocal regulation of ion-channel activity, membrane potential, and the function of receptor complexes may contribute to the proper functioning of the immunological synapse.