Conjugated polythiophene probes target lysosome-related acidic vacuoles in cultured primary cells
Conjugated polythiophene probes target lysosome-related acidic vacuoles in cultured primary cells
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DOI：
10.1016/j.mcp.2007.04.005
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发表时间：
2007-10-01
影响因子：
3.3
通讯作者：
Jonasson, Jon
中科院分区：
文献类型：
--
作者：
Bjork, Per;Nilsson, K. Peter R.;Jonasson, Jon
用于研究生物过程和结构的构象敏感的光学探针是非常感兴趣的。目前的工作表明，共轭的CPE（CPE）探针可用于特异性靶向染色质，核和胞质囊泡，和细胞骨架成分在一个复杂的系统培养的细胞。其中一种探针也可用于活细胞的活体染色。当与不同的实体结合时，聚噻吩衍生物探针由于这些构象敏感探针的独特光谱性质而发射具有不同颜色的光。结合的物理预处理也可以用于表征靶标。出乎意料的是，溶酶体相关的酸性空泡培养的原代细胞的阴离子，阳离子和两性离子聚噻吩衍生物的目标。用Bafilomycin At（一种液泡型H+-ATP酶的特异性抑制剂）预处理引起染色的重新分布。在转化细胞（黑色素瘤、神经母细胞瘤和前列腺癌细胞系）中无法证明溶酶体相关酸性囊泡的靶向作用，这表明与恶性细胞系相比，培养的正常细胞中靶点的定位、结构、可及性或数量存在差异。在细胞质空泡中的共轭复合物的化学性质仍然难以捉摸。(c)2007爱思唯尔有限公司保留所有权利。
Conformation-sensitive optical probes for studying biological processes and structures are of great interest. The present work shows for the first time that conjugated polyelectrolyte (CPE) probes can be used for specific targeting of chromatin, nuclear and cytoplasmatic vesicles, and cytoskeletal components in a complex system of cultured cells. One of the probes could also be used for vital staining of live cells. When bound to different entities, the polythiophene derivative probes emitted light with different colors due to the unique spectral properties of these conformation sensitive probes. The physical pre-requisites for binding could also be exploited for characterization of the target. Unexpectedly, lysosome-related acidic vacuoles were targeted in cultured primary cells by both anionic, cationic, and zwitterionic polythiophene derivatives. Pre-treatment with Bafilomycin At, a specific inhibitor of vacuolar-type H+-ATPase, caused redistribution of the staining. The targeting of lysosome-related acidic vesicles could not be demonstrated in transformed cells (melanoma, neuroblastoma, and prostate cancer cell lines), indicating a difference in the localization, structure, accessibility, or quantity of the target in cultured normal cells as compared with the malignant cell lines. The chemical nature of the conjugated polyelectrolyte complex in the cytoplasmatic vacuoles remains elusive. (c) 2007 Elsevier Ltd. All rights reserved.