Björk Per, Peter R Nilsson K, Lenner Liselotte, Kågedal Bertil, Persson Birgitta, Inganäs Olle, Jonasson Jon
Biomolecular and Organic Electronics, IFM, Linköping University, SE-581 83 Linköping, Sweden.
Mol Cell Probes. 2007 Oct-Dec;21(5-6):329-37. doi: 10.1016/j.mcp.2007.04.005. Epub 2007 Apr 27.
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 zwitter-ionic polythiophene derivatives. Pre-treatment with Bafilomycin A1, 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.
用于研究生物过程和结构的构象敏感光学探针备受关注。目前的工作首次表明,共轭聚电解质(CPE)探针可用于在培养细胞的复杂系统中特异性靶向染色质、核和细胞质囊泡以及细胞骨架成分。其中一种探针还可用于活细胞的活体染色。当与不同实体结合时,由于这些构象敏感探针的独特光谱特性,聚噻吩衍生物探针会发出不同颜色的光。结合的物理先决条件也可用于表征靶标。出乎意料的是,阴离子、阳离子和两性离子聚噻吩衍生物在培养的原代细胞中均靶向溶酶体相关酸性液泡。用液泡型H(+)-ATP酶的特异性抑制剂巴弗洛霉素A1预处理会导致染色重新分布。在转化细胞(黑色素瘤、神经母细胞瘤和前列腺癌细胞系)中未证实溶酶体相关酸性囊泡的靶向,这表明与恶性细胞系相比,培养的正常细胞中靶标的定位、结构、可及性或数量存在差异。细胞质液泡中共轭聚电解质复合物的化学性质仍然难以捉摸。