Department of Physics, Boise State University, Boise, ID, USA.
Department of Biology, Boise State University, Boise, ID, USA.
Biochim Biophys Acta Biomembr. 2021 Aug 1;1863(8):183625. doi: 10.1016/j.bbamem.2021.183625. Epub 2021 Apr 20.
Cholesterol (Chol) content in most cellular membranes does not exceed 50 mol%, only in the eye lens's fiber cell plasma membrane, its content surpasses 50 mol%. At this high concentration, Chol induces the formation of pure cholesterol bilayer domains (CBDs), which coexist with the surrounding phospholipid-cholesterol domain (PCD). Here, we applied atomic force microscopy to study the mechanical properties of Chol/phosphatidylcholine membranes where the Chol content was increased from 0 to 75 mol%, relevant to eye lens membranes. The surface roughness of the membrane decreases with an increase of Chol content until it reaches 60 mol%, and roughness increases with a further increment in Chol content. We propose that the increased roughness at higher Chol content results from the formation of CBDs. Force spectroscopy on the membrane with Chol content of 50 mol% or lesser exhibited single breakthrough events, whereas two distinct puncture events were observed for membranes with the Chol content greater than 50 mol%. We propose that the first puncture force corresponds to the membranes containing coexisting PCD and CBDs. In contrast, the second puncture force corresponds to the "CBD water pocket" formed due to coexisting CBDs and PCD. Membrane area compressibility modulus (K) increases with an increase in Chol content until it reaches 60 mol%, and with further increment in Chol content, CBDs are formed, and K starts to decrease. Our results report the increase in membrane roughness and decrease K at very high Chol content (>60 mol%) relevant to the eye lens membrane.
胆固醇(Chol)在大多数细胞膜中的含量不超过 50mol%,仅在眼睛晶状体的纤维细胞膜中,其含量超过 50mol%。在这种高浓度下,Chol 诱导形成纯胆固醇双层域(CBD),与周围的磷脂-胆固醇域(PCD)共存。在这里,我们应用原子力显微镜研究胆固醇/磷脂膜的机械性能,胆固醇含量从 0 增加到 75mol%,与眼睛晶状体膜相关。随着胆固醇含量的增加,膜的表面粗糙度降低,直到达到 60mol%,然后随着胆固醇含量的进一步增加而增加。我们提出,在较高胆固醇含量下粗糙度的增加是由于 CBD 的形成。胆固醇含量为 50mol%或更低的膜上的力谱分析显示出单个突破事件,而胆固醇含量大于 50mol%的膜上观察到两个明显的穿孔事件。我们提出,第一次穿刺力对应于含有共存 PCD 和 CBD 的膜。相比之下,第二次穿刺力对应于由于共存 CBD 和 PCD 形成的“CBD 水口袋”。膜面积压缩模量(K)随着胆固醇含量的增加而增加,直到达到 60mol%,随着胆固醇含量的进一步增加,形成了 CBD,K 开始下降。我们的结果报告了与眼睛晶状体膜相关的非常高的胆固醇含量(>60mol%)下膜粗糙度的增加和 K 的降低。