Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA.
Bioconjug Chem. 2011 Nov 16;22(11):2345-54. doi: 10.1021/bc200424r. Epub 2011 Oct 28.
Fluorescence quenching groups are widely employed in biological detection, sensing, and imaging. To date, a relatively small number of such groups are in common use. Perhaps the most commonly used quencher, dabcyl, has limited efficiency with a broad range of fluorophores. Here, we describe a molecular approach to improve the efficiency of quenchers by increasing their electronic complexity. Multi-Path Quenchers (MPQ) are designed to have multiple donor or acceptor groups in their structure, allowing for a multiplicity of conjugation pathways of varied length. This has the effect of broadening the absorption spectrum, which in turn can increase quenching efficiency and versatility. Six such MPQ derivatives are synthesized and tested for quenching efficiency in a DNA hybridization context. Duplexes placing quenchers and fluorophores within contact distance or beyond this distance are used to measure quenching via contact or FRET mechanisms. Results show that several of the quenchers are considerably more efficient than dabcyl at quenching a wider range of common fluorophores, and two quench fluorescein and TAMRA as well as or better than a Black Hole Quencher.
荧光猝灭基团广泛应用于生物检测、传感和成像。迄今为止，这类基团的应用相对较少。也许最常用的猝灭剂 dabcyl 的效率有限，且适用于广泛的荧光团。在这里，我们描述了一种通过增加电子复杂性来提高猝灭剂效率的分子方法。多路径猝灭剂 (MPQ) 的设计结构中具有多个供体或受体基团，允许存在多种不同长度的共轭途径。这会拓宽吸收光谱，从而可以提高猝灭效率和多功能性。合成了六个这样的 MPQ 衍生物，并在 DNA 杂交背景下测试其猝灭效率。将猝灭剂和荧光团置于接触距离内或之外的双链体用于通过接触或 FRET 机制测量猝灭。结果表明，与 dabcyl 相比，几种猝灭剂在猝灭更广泛范围的常见荧光团方面效率更高，并且有两个猝灭剂可以与 Black Hole Quencher 一样或更好地猝灭荧光素和 TAMRA。
