Bandichhor Rakeshwar, Petrescu Anca D, Vespa Aude, Kier Ann B, Schroeder Friedhelm, Burgess Kevin
Department of Chemistry, Texas A and M University, Box 30012, College Station, Texas 77841, USA.
J Am Chem Soc. 2006 Aug 23;128(33):10688-9. doi: 10.1021/ja063784a.
A special, water-soluble, fluorescent probe 1 was designed. This consisted of a fluorescein-based component to harvest irradiation at 488 nm and a rhodamine-based part designed to emit it at a significantly longer wavelength. This cassette was used to label an illustrative protein called ACBP. Evidence was accumulated to support the assertion that ACBP-1 bound its native ligand with a binding constant similar to that of the unlabeled protein, and retained its secondary structure (CD). ACBP-1 was imported into cells using the Chariot peptide. Confocal images proved that some ACBP-1 localized into the nucleus (as expected) and, most significantly, it could be visualized more effectively by irradiating at the donor (fluorescein-like) part of the cassette, than the acceptor (rhodamine-like) part. Overall, this study demonstrates that cassettes of this kind can label a protein without significantly perturbing its function or secondary structure and they can be visualized effectively via irradiation of the donor and observation of the acceptor fluorescence.
设计了一种特殊的水溶性荧光探针1。它由一个基于荧光素的成分组成，用于收集488nm的辐射，以及一个基于罗丹明的部分，设计用于在显著更长的波长处发射辐射。该盒式结构用于标记一种名为ACBP的示例性蛋白质。积累的证据支持以下论断：ACBP-1与其天然配体结合，结合常数与未标记的蛋白质相似，并保留其二级结构（圆二色性）。使用Chariot肽将ACBP-1导入细胞。共聚焦图像证明，一些ACBP-1定位到细胞核中（如预期），最重要的是，通过照射盒式结构的供体（类似荧光素）部分，比受体（类似罗丹明）部分，能更有效地观察到它。总体而言，这项研究表明，这种盒式结构可以标记蛋白质，而不会显著干扰其功能或二级结构，并且可以通过照射供体并观察受体荧光来有效地进行可视化。
