Department of Chemistry, University of Washington, Seattle, Washington 98195, United States.
J Phys Chem B. 2011 Aug 4;115(30):9452-60. doi: 10.1021/jp1101829. Epub 2011 Jul 11.
LKα14 is a 14 amino acid peptide with a periodic sequence of leucine and lysine residues consistent with an amphipathic α-helix. This "hydrophobic periodicity" has been found to result in an α-helical secondary structure at air-water interfaces and on both polar and nonpolar solid polymer surfaces. In this paper, the dynamics of LKα14 peptides, selectively deuterated at a single leucine and adsorbed onto polystyrene and carboxylated polystyrene beads, are studied using (2)H magic angle spinning (MAS) solid state NMR over a 100 °C temperature range. We first demonstrate the sensitivity enhancement possible with (2)H MAS techniques, which in turn enables us to obtain high-quality (2)H NMR spectra for selectively deuterated peptides adsorbed onto solid polymer surfaces. The extensive literature shows that the dynamics of leucine side chains are sensitive to the local structural environment of the protein. Therefore, the degree to which the dynamics of leucine side chains and the backbone of the peptide LKα14 are influenced by surface proximity and surface chemistry is studied as a function of temperature with (2)H MAS NMR. It is found that the dynamics of the leucine side chains in LKα14 depend strongly upon the orientation of the polymer on the surface, which in turn depends on whether the LKα14 peptide adsorbs onto a polar or nonpolar surface. (2)H MAS line shapes therefore permit probes of surface orientation over a wide temperature range.
LKα14 是一种由 14 个氨基酸组成的肽,其亮氨酸和赖氨酸残基呈周期性排列,符合两亲性α-螺旋结构。这种“疏水性周期性”导致其在气-水界面以及极性和非极性聚合物表面上形成α-螺旋二级结构。在本文中,我们使用(2)H 魔角旋转(MAS)固态 NMR 技术,在 100°C 的温度范围内,研究了在单个亮氨酸处选择性氘代并吸附在聚苯乙烯和羧基化聚苯乙烯珠上的 LKα14 肽的动力学。我们首先证明了(2)H MAS 技术的灵敏度增强效果,这反过来使我们能够为吸附在固体聚合物表面上的选择性氘代肽获得高质量的(2)H NMR 谱。大量文献表明,亮氨酸侧链的动力学对蛋白质的局部结构环境敏感。因此,我们研究了肽 LKα14 的亮氨酸侧链和肽骨架的动力学受表面接近和表面化学的影响程度随温度的变化。结果发现,LKα14 中亮氨酸侧链的动力学强烈依赖于表面聚合物的取向,而这又取决于 LKα14 肽吸附到极性还是非极性表面上。因此,(2)H MAS 谱线形状允许在较宽的温度范围内探测表面取向。