Department of Biophysics, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA.
Exp Eye Res. 2012 Apr;97(1):117-29. doi: 10.1016/j.exer.2012.01.012. Epub 2012 Feb 2.
The organization and physical properties of the lipid bilayer portion of intact cortical and nuclear fiber cell plasma membranes isolated from the eye lenses of two-year-old pigs were studied using electron paramagnetic resonance (EPR) spin-labeling. Membrane fluidity, hydrophobicity, and the oxygen transport parameter (OTP) were assessed from the EPR spectra of precisely positioned spin labels. Intact cortical and nuclear membranes, which include membrane proteins, were found to contain three distinct lipid environments. These lipid environments were termed the bulk lipid domain, boundary lipid domain, and trapped lipid domain (lipids in protein aggregates). The amount of boundary and trapped lipids was greater in intact nuclear membranes than in cortical membranes. The properties of intact membranes were compared with the organization and properties of lens lipid membranes made of the total lipid extracts from the lens cortex or nucleus. In cortical lens lipid membranes, only one homogenous environment was detected, which was designated as a bulk lipid domain (phospholipid bilayer saturated with cholesterol). Lens lipid membranes prepared from the lens nucleus possessed two domains, assigned as a bulk lipid domain and a cholesterol bilayer domain (CBD). In intact nuclear membranes, it was difficult to discriminate the CBD, which was clearly detected in nuclear lens lipid membranes, because the OTP measured in the CBD is the same as in the domain formed by trapped lipids. The two domains unique to intact membranes-namely, the domain formed by boundary lipids and the domain formed by trapped lipids-were most likely formed due to the presence of membrane proteins. It is concluded that formation of rigid and practically impermeable domains is enhanced in the lens nucleus, indicating changes in membrane composition that may help to maintain low oxygen concentration in this lens region.
我们使用电子顺磁共振(EPR)自旋标记研究了从两年龄猪晶状体中分离出的完整皮质和核纤维细胞质膜的脂质双层部分的组织和物理性质。从精确定位的自旋标记的 EPR 谱评估了膜流动性、疏水性和氧传递参数(OTP)。完整的皮质和核膜,其中包括膜蛋白,被发现包含三个不同的脂质环境。这些脂质环境分别称为体相脂质域、边界脂质域和束缚脂质域(蛋白聚集体中的脂质)。完整核膜中边界和束缚脂质的量大于皮质膜。将完整膜的性质与由晶状体皮质或核的总脂质提取物制成的晶状体脂质膜的组织和性质进行了比较。在皮质晶状体脂质膜中,仅检测到一种均匀的环境,被指定为体相脂质域(饱和胆固醇的磷脂双层)。从晶状体核制备的晶状体脂质膜具有两个域,分别指定为体相脂质域和胆固醇双层域(CBD)。在完整的核膜中,由于 CBD 中测量的 OTP 与束缚脂质形成的域中的 OTP 相同,因此很难区分 CBD,而在核晶状体脂质膜中 CBD 则清晰可辨。这两个仅存在于完整膜中的域——即边界脂质形成的域和束缚脂质形成的域——很可能是由于膜蛋白的存在而形成的。研究结论认为,晶状体核中刚性和几乎不可渗透的域的形成得到了增强,这表明膜组成发生了变化,这可能有助于维持该晶状体区域的低氧浓度。