Yun C S, Javier A, Jennings T, Fisher M, Hira S, Peterson S, Hopkins B, Reich N O, Strouse G F
Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida 32306-4390, USA.
J Am Chem Soc. 2005 Mar 9;127(9):3115-9. doi: 10.1021/ja043940i.
Optical-based distance measurements are essential for tracking biomolecular conformational changes, drug discovery, and cell biology. Traditional Forster resonance energy transfer (FRET) is efficient for separation distances up to 100 A. We report the first successful application of a dipole-surface type energy transfer from a molecular dipole to a nanometal surface that more than doubles the traditional Forster range (220 A) and follows a 1/R(4) distance dependence. We appended a 1.4 nm Au cluster to the 5' end of one DNA strand as the energy acceptor and a fluorescein (FAM) to the 5' end of the complementary strand as the energy donor. Analysis of the energy transfer on DNA lengths (15, 20, 30, 60bp), complemented by protein-induced DNA bending, provides the basis for fully mapping the extent of this dipole surface type mechanism over its entire usable range (50-250 A). Further, protein function is fully compatible with these nanometal-DNA constructs. Significantly extending the range of optical based methods in molecular rulers is an important leap forward for biophysics.
基于光学的距离测量对于追踪生物分子构象变化、药物发现和细胞生物学至关重要。传统的福斯特共振能量转移(FRET)对于高达100埃的分离距离是有效的。我们报道了首次成功应用从分子偶极子到纳米金属表面的偶极-表面型能量转移,其使传统的福斯特范围(220埃)增加了一倍多,并遵循1/R(4)距离依赖性。我们在一条DNA链的5'端附加了一个1.4纳米的金簇作为能量受体,并在互补链的5'端附加了一个荧光素(FAM)作为能量供体。对DNA长度(15、20、30、60碱基对)上的能量转移进行分析,并辅以蛋白质诱导的DNA弯曲,为在其整个可用范围(50-250埃)内全面绘制这种偶极表面型机制的范围提供了基础。此外,蛋白质功能与这些纳米金属-DNA构建体完全兼容。显著扩展分子尺中基于光学方法的范围是生物物理学向前迈出的重要一步。