Peterlin Primoz, Arrigler Vesna, Kogej Ksenija, Svetina Sasa, Walde Peter
University of Ljubljana, Faculty of Medicine, Institute of Biophysics, Lipiceva 2, SI-1000 Ljubljana, Slovenia.
Chem Phys Lipids. 2009 Jun;159(2):67-76. doi: 10.1016/j.chemphyslip.2009.03.005. Epub 2009 Apr 2.
The interaction of two types of vesicle systems was investigated: micrometer-sized, giant unilamellar vesicles (GUVs) formed from 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and submicrometer-sized, large unilamellar vesicles (LUVs) formed from oleic acid and oleate, both in a buffered aqueous solution (pH 8.8). Individual POPC GUVs were transferred with a micropipette into a suspension of oleic acid/oleate LUVs, and the shape changes of the GUVs were monitored using optical microscopy. The behavior of POPC GUVs upon transfer into a 0.8mM suspension of oleic acid, in which oleic acid/oleate forms vesicular bilayer structures, was qualitatively different from the behavior upon transfer into a 0.3mM suspension of oleic acid/oleate, in which oleic acid/oleate is predominantly present in the form of monomers and possibly non-vesicular aggregates. In both cases, changes in vesicle morphology were observed within tens of seconds after the transfer. After an initial increase of the vesicle cross-section, the vesicle started to evaginate, spawning dozens of satellite vesicles connected to the mother vesicle with narrow necks or tethers. In 60% of the cases of transfer into a 0.8mM oleic acid suspension, the evagination process reversed and proceeded to the point where the membrane formed invaginations. In some of these cases, several consecutive transitions between invaginated and evaginated shapes were observed. In the remaining 40% of the cases of transfer into the 0.8mM oleic acid suspension and in all cases of vesicle transfer into the 0.3mM oleic acid suspension, no invaginations nor subsequent evaginations were observed. An interpretation of the observed vesicle shape transformation on the basis of the bilayer-couple model is proposed, which takes into account uptake of oleic acid/oleate molecules by the POPC vesicles, oleic acid flip-flop processes and transient pore formation.
由1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱(POPC)形成的微米级巨型单层囊泡(GUVs),以及由油酸和油酸盐在缓冲水溶液(pH 8.8)中形成的亚微米级大单层囊泡(LUVs)。用微量移液器将单个POPC GUV转移到油酸/油酸盐LUV的悬浮液中,并使用光学显微镜监测GUV的形状变化。将POPC GUV转移到0.8mM油酸悬浮液(其中油酸/油酸盐形成囊泡双层结构)时的行为,与转移到0.3mM油酸/油酸盐悬浮液(其中油酸/油酸盐主要以单体形式存在,可能还有非囊泡聚集体)时的行为在性质上有所不同。在这两种情况下,转移后几十秒内都观察到了囊泡形态的变化。在囊泡横截面最初增加后,囊泡开始外翻,产生数十个通过窄颈或系链与母囊泡相连的卫星囊泡。在转移到0.8mM油酸悬浮液的60%的情况下,外翻过程逆转并进行到膜形成内陷的程度。在其中一些情况下,观察到内陷和外翻形状之间的几次连续转变。在转移到0.8mM油酸悬浮液的其余40%的情况下,以及在所有将囊泡转移到0.3mM油酸悬浮液的情况下,均未观察到内陷和随后的外翻。基于双层偶合模型对观察到的囊泡形状转变提出了一种解释,该解释考虑了POPC囊泡对油酸/油酸盐分子的摄取、油酸的翻转过程和瞬时孔的形成。