School of Physics and Astronomy, University of Leeds, Leeds LS2 9JT, UK.
J Phys Condens Matter. 2010 Jun 16;22(23):235103. doi: 10.1088/0953-8984/22/23/235103. Epub 2010 May 21.
Fluorescence resonance energy transfer is a powerful technique which is often used to probe the properties of proteins and complex macromolecules. The technique relies on relatively large fluorescent dyes which are engineered into the molecule of interest. In the case of small proteins, these dyes may affect the stability of the protein, and modify the folding kinetics and the folding mechanisms which are being probed. Here we use atomistic simulation to investigate the effect that commonly used fluorescent dyes have on the folding of a four-helix bundle protein. We show that, depending on where the dyes are attached, their effect on the kinetic and thermodynamic properties of the protein may be significant. We find that, while the overall folding mechanism is not affected by the dyes, they can destabilize, or even stabilize, intermediate states.
荧光共振能量转移是一种强大的技术，常用于探测蛋白质和复杂大分子的性质。该技术依赖于相对较大的荧光染料，这些染料被设计到感兴趣的分子中。在小蛋白质的情况下，这些染料可能会影响蛋白质的稳定性，并改变正在探测的折叠动力学和折叠机制。在这里，我们使用原子模拟来研究常用荧光染料对四螺旋束蛋白质折叠的影响。我们表明，根据染料附着的位置，它们对蛋白质动力学和热力学性质的影响可能是显著的。我们发现，虽然染料不会影响整体折叠机制，但它们可以使中间状态失稳，甚至稳定。
