Graduate Program in Biophysics, University of Wisconsin, Madison, 53706, United States.
Biochemistry. 2011 Jan 18;50(2):266-75. doi: 10.1021/bi101096k. Epub 2010 Dec 15.
Revealing the thermodynamic driving force of protein-DNA interactions is crucial to the understanding of factors that dictate the properties and function of protein-DNA complexes. For the binding of DNA to DNA-wrapping proteins, such as the integration host factor (IHF), Record and co-workers proposed that the disruption of a large number of preexisting salt bridges is coupled with the binding process [Holbrook, J. A., et al. (2001) J. Mol. Biol. 310, 379]. To test this proposal, we have conducted explicit solvent MD simulations (multiple ∼25-50 ns trajectories for each salt concentration) to examine the behavior of charged residues in IHF, especially concerning their ability to form salt bridges at different salt concentrations. Of the 17 cationic residues noted by Record and co-workers, most are engaged in salt bridge interactions for a significant portion of the trajectories, especially in the absence of salt. This observation suggests that, from a structural point of view, their proposal is plausible. However, the complex behaviors of charged residues observed in the MD simulations also suggest that the unusual thermodynamic characteristics of IHF-DNA binding likely arise from the interplay between complex dynamics of charged residues both in and beyond the DNA binding site. Moreover, a comparison of MD simulations at different salt concentrations suggests that the strong dependence of the IHF-DNA binding enthalpy on salt concentration may not be due to a significant decrease in the number of stable salt bridges in apo IHF at high salt concentrations. In addition to the Hofmeister effects quantified in more recent studies of IHF-DNA binding, we recommend consideration of the variation of the enthalpy change of salt bridge disruption at different salt concentrations. Finally, the simulation study presented here explicitly highlights the fact that the electrostatic properties of DNA-binding proteins can be rather different in the apo and DNA-bound states, which has important implications for the design of robust methods for predicting DNA binding sites in proteins.
揭示蛋白质 - DNA 相互作用的热力学驱动力对于理解决定蛋白质 - DNA 复合物性质和功能的因素至关重要。对于 DNA 与 DNA 包装蛋白(如整合宿主因子(IHF))的结合,Record 及其同事提出,破坏大量预先存在的盐桥与结合过程相关联 [Holbrook, J. A., et al. (2001) J. Mol. Biol. 310, 379]。为了验证这一假设,我们进行了显式溶剂 MD 模拟(每种盐浓度下进行多个 ∼25-50 ns 的轨迹),以研究 IHF 中带电残基的行为,特别是它们在不同盐浓度下形成盐桥的能力。Record 及其同事指出的 17 个阳离子残基中,大多数在很大一部分轨迹中参与盐桥相互作用,尤其是在没有盐的情况下。这一观察结果表明,从结构角度来看,他们的假设是合理的。然而,MD 模拟中观察到的带电残基的复杂行为也表明,IHF-DNA 结合的异常热力学特征可能源于 DNA 结合位点内外带电残基的复杂动力学相互作用。此外,不同盐浓度下的 MD 模拟比较表明,IHF-DNA 结合焓对盐浓度的强烈依赖性可能不是由于在高盐浓度下 apo IHF 中稳定盐桥数量的显著减少所致。除了最近对 IHF-DNA 结合研究中量化的 Hofmeister 效应外,我们建议考虑不同盐浓度下盐桥破坏焓变的变化。最后,这里提出的模拟研究明确强调了一个事实,即 DNA 结合蛋白的静电特性在 apo 和 DNA 结合状态下可能有很大的不同,这对设计预测蛋白质中 DNA 结合位点的稳健方法具有重要意义。