Nakahara Hiromichi, Lee Sannamu, Shibata Osamu
Department of Biophysical Chemistry, Faculty of Pharmaceutical Sciences, Nagasaki International University, Nagasaki, Japan.
Biophys J. 2009 Feb 18;96(4):1415-29. doi: 10.1016/j.bpj.2008.11.022.
Interfacial behavior was studied in pulmonary surfactant model systems containing an amphiphilic alpha-helical peptide (Hel 13-5), which consists of 13 hydrophobic and five hydrophilic amino acid residues. Fully saturated phospholipids of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) were utilized to understand specific interactions between anionic DPPG and cationic Hel 13-5 for pulmonary functions. Surface pressure (pi)-molecular area (A) and surface potential (DeltaV)-A isotherms of DPPG/Hel 13-5 and DPPC/DPPG (4:1, mol/mol)/Hel 13-5 preparations were measured to obtain basic information on the phase behavior under compression and expansion processes. The interaction leads to a variation in squeeze-out surface pressures against a mole fraction of Hel 13-5, where Hel 13-5 is eliminated from the surface on compression. The phase behavior was visualized by means of Brewster angle microscopy, fluorescence microscopy, and atomic force microscopy. At low surface pressures, the formation of differently ordered domains in size and shape is induced by electrostatic interactions. The domains independently grow upon compression to high surface pressures, especially in the DPPG/Hel 13-5 system. Under the further compression process, protrusion masses are formed in AFM images in the vicinity of squeeze-out pressures. The protrusion masses, which are attributed to the squeezed-out Hel 13-5, grow larger in lateral size with increasing DPPG content in phospholipid compositions. During subsequent expansion up to 35 mN m(-1), the protrusions retain their height and lateral diameter for the DPPG/Hel 13-5 system, whereas the protrusions become smaller for the DPPC/Hel 13-5 and DPPC/DPPG/Hel 13-5 systems due to a reentrance of the ejected Hel 13-5 into the surface. In this work we detected for the first time, to our knowledge, a remarkably large hysteresis loop for cyclic DeltaV-A isotherms of the binary DPPG/Hel 13-5 preparation. This exciting phenomenon suggests that the specific interaction triggers two completely independent processes for Hel 13-5 during repeated compression and expansion: 1), squeezing-out into the subsolution; and 2), and close packing as a monolayer with DPPG at the interface. These characteristic processes are also strongly supported by atomic force microscopy observations. The data presented here provide complementary information on the mechanism and importance of the specific interaction between the phosphatidylglycerol headgroup and the polarized moiety of native surfactant protein B for biophysical functions of pulmonary surfactants.
在含有两亲性α-螺旋肽(Hel 13-5)的肺表面活性剂模型系统中研究了界面行为,该肽由13个疏水氨基酸残基和5个亲水氨基酸残基组成。使用二棕榈酰磷脂酰胆碱(DPPC)和二棕榈酰磷脂酰甘油(DPPG)的完全饱和磷脂来了解阴离子DPPG和阳离子Hel 13-5之间对于肺功能的特定相互作用。测量了DPPG/Hel 13-5和DPPC/DPPG(4:1,摩尔/摩尔)/Hel 13-5制剂的表面压力(π)-分子面积(A)和表面电位(ΔV)-A等温线,以获得关于压缩和膨胀过程中相行为的基本信息。这种相互作用导致针对Hel 13-5摩尔分数的挤出表面压力发生变化,其中Hel 13-5在压缩时从表面被消除。通过布鲁斯特角显微镜、荧光显微镜和原子力显微镜观察相行为。在低表面压力下,静电相互作用诱导形成大小和形状不同的有序域。这些域在压缩至高表面压力时独立生长,特别是在DPPG/Hel 13-5系统中。在进一步的压缩过程中,在AFM图像中挤出压力附近形成突出物。这些突出物归因于挤出的Hel 13-5,随着磷脂组合物中DPPG含量的增加,其横向尺寸变得更大。在随后膨胀至35 mN m-1的过程中,对于DPPG/Hel 13-5系统,突出物保持其高度和横向直径,而对于DPPC/Hel 13-5和DPPC/DPPG/Hel 13-5系统,由于喷出的Hel 13-5重新进入表面,突出物变得更小。据我们所知,在这项工作中我们首次检测到二元DPPG/Hel 13-5制剂的循环ΔV-A等温线存在非常大的滞后环。这一令人兴奋的现象表明,这种特定相互作用在重复压缩和膨胀过程中触发了Hel 13-5的两个完全独立的过程:1),挤出到亚溶液中;以及2),作为单层与界面处的DPPG紧密堆积。原子力显微镜观察也有力地支持了这些特征过程。此处呈现的数据为磷脂酰甘油头部基团与天然表面活性剂蛋白B的极化部分之间特定相互作用对于肺表面活性剂生物物理功能的机制和重要性提供了补充信息。