Zweckstetter Markus
Department of NMR-based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
Eur Biophys J. 2006 Jan;35(2):170-80. doi: 10.1007/s00249-005-0018-6. Epub 2005 Oct 26.
Recently we reported that the alignment tensor of a biological macromolecule, which was dissolved in a dilute suspension of highly negatively charged filamentous phage at close to neutral pH, can be predicted from the molecule's 3D charge distribution and shape (Zweckstetter et al. 2004). Here it is demonstrated that this approach is also applicable to alignment of proteins in liquid crystalline phases formed by filamentous phage at low pH. Residual dipolar couplings (RDCs) predicted by our simple electrostatic model for the B1 domain of protein G in fd phage at pH 3 fit very well with the experimental values. The sign of charge-shape predicted one-bond (1)H-(15)N dipolar couplings for the B1 domain of protein G (GB1) was inverted at pH 3 compared to neutral pH, in agreement with experimental observations. Our predictions indicate that this is a feature specific for GB1. In addition, it is shown that RDCs induced in the protein ubiquitin by the presence of a positively charged surfactant system comprising cetylpyridinium bromide/hexanol/sodium bromide can be predicted accurately by a simple electrostatic alignment model. This shows that steric and electrostatic interactions dominate weak alignment of biomolecules for a wide range of pH values both in filamentous phage and in surfactant liquid crystalline phases.
最近我们报道,在接近中性pH值的条件下,溶解于带高负电荷丝状噬菌体的稀悬浮液中的生物大分子的取向张量,可以根据分子的三维电荷分布和形状来预测(茨韦克施泰特等人,2004年)。本文证明,这种方法也适用于在低pH值下由丝状噬菌体形成的液晶相中蛋白质的取向。我们的简单静电模型预测的pH值为3时fd噬菌体中蛋白G的B1结构域的剩余偶极耦合(RDCs)与实验值非常吻合。与中性pH值相比,蛋白G(GB1)的B1结构域的电荷-形状预测的一键(1)H-(15)N偶极耦合在pH值为3时符号反转,这与实验观察结果一致。我们的预测表明,这是GB1特有的特征。此外,研究表明,由包含十六烷基溴化吡啶/己醇/溴化钠的带正电荷表面活性剂体系在蛋白质泛素中诱导产生的RDCs,可以通过简单的静电取向模型准确预测。这表明,在丝状噬菌体和表面活性剂液晶相中,对于广泛的pH值范围,空间和静电相互作用主导着生物分子的弱取向。