Department of Otorhinolaryngology, Head and Neck Surgery, Section for Experimental Oncology and Nanomedicine (SEON), University Hospital, Waldstraße 1/Glückstraße 10, D-91054 Erlangen, Germany. Department for Internal Medicine 3, University Hospital Erlangen, Friedrich-Alexander University of Erlangen-Nuremberg, Ulmenweg 18, D-91054 Erlangen, Germany.
Phys Biol. 2013 Dec;10(6):065006. doi: 10.1088/1478-3975/10/6/065006. Epub 2013 Dec 4.
Healthy cells exhibit an asymmetric plasma membrane with phosphatidylserine (PS) located on the cytoplasmic leaflet of the plasma membrane bilayer. Annexin A5-FITC, a PS binding protein, is commonly used to evaluate apoptosis in flow cytometry. PS exposed by apoptotic cells serves as a major 'eat-me' signal for phagocytes. Although exposition of PS has been observed after alternative stimuli, no clearance of viable, PS exposing cells has been detected. Thus, besides PS exposure, membranes of viable and apoptotic cells might exhibit specific characteristics. Here, we show that Annexin A5 binds in a cooperative manner to different types of dead cells. Shrunken apoptotic cells thereby showed the highest Hill coefficient values. Contrarily, parafomaldehyde fixation of apoptotic cells completely abrogates the cooperativity effect seen with dead and dying cells. We tend to speculate that the cooperative binding of Annexin A5 to the membranes of apoptotic cells reflects higher fluidity of the exposed membranes facilitating PS clustering.
健康细胞表现出不对称的质膜,其质膜双层的细胞质小叶中存在磷脂酰丝氨酸 (PS)。膜联蛋白 A5-FITC 是一种 PS 结合蛋白,常用于流式细胞术评估细胞凋亡。凋亡细胞暴露的 PS 作为吞噬细胞的主要“吃我”信号。尽管在替代刺激后观察到 PS 的暴露,但未检测到有活力、暴露 PS 的细胞的清除。因此,除 PS 暴露外,活细胞和凋亡细胞的膜可能表现出特定的特征。在这里,我们表明膜联蛋白 A5 以协同的方式结合到不同类型的死亡细胞上。因此,皱缩的凋亡细胞显示出最高的 Hill 系数值。相反,用多聚甲醛固定凋亡细胞会完全消除与死亡和垂死细胞一起观察到的协同作用。我们倾向于推测膜联蛋白 A5 与凋亡细胞的膜的协同结合反映了暴露膜的更高流动性,从而促进 PS 聚集。