Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, China.
Analyst. 2013 Feb 21;138(3):825-30. doi: 10.1039/c2an36322d. Epub 2012 Dec 14.
A new spectrofluorometric method for cellular prion protein (PrP(C)) was developed based on the regulation of N,N'-bis[3,3'-(dimethylamino)propylamine]-3,4,9,10-perylenetetracarboxylic diimide (DAPER) fluorescence. As a perylene derivative, DAPER emits strong fluorescence in the form of free monomer in aqueous medium, but not in the form of aggregates. In this contribution, we found that the aptamer of PrP(C) could induce the aggregation of DAPER, and the bright fluorescence of DAPER was completely quenched. The quenched fluorescence, however, was recovered if PrP(C) was further added, which was ascribed to the specific binding of PrP(C) to its aptamer and the releasing of free DAPER monomers. This signalling mechanism makes it possible to detect PrP(C) by fluorescence spectroscopy. The assay allows the selective determination of PrP(C) in aqueous solution with high sensitivity and exhibits a good linear range from 0.4 to 1.6 nmol L(-1). Moreover, this probe can be applied to monitor the level of PrP(C) in human urine samples with satisfactory results.
基于 N,N'-双[3,3'-(二甲氨基)丙基]胺]-3,4,9,10-苝四羧酸二酰亚胺(DAPER)荧光调控,建立了一种新的细胞朊蛋白(PrP(C))分光荧光测定法。作为一种苝衍生物,DAPER 在水介质中以游离单体的形式发出强荧光,但不以聚集体的形式发出荧光。在本研究中,我们发现 PrP(C) 的适体能够诱导 DAPER 的聚集,从而使 DAPER 的强荧光完全猝灭。然而,如果进一步加入 PrP(C),猝灭的荧光会恢复,这归因于 PrP(C)与其适体的特异性结合以及游离 DAPER 单体的释放。这种信号机制使得通过荧光光谱法检测 PrP(C)成为可能。该测定法允许在水溶液中以高灵敏度选择性地测定 PrP(C),并表现出从 0.4 到 1.6 nmol L(-1) 的良好线性范围。此外,该探针可用于监测人尿样中 PrP(C)的水平,结果令人满意。