Properties of fiber cell plasma membranes isolated from the cortex and nucleus of the porcine eye lens

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• 作者：Laxman Mainali^a ; ¹ ; Marija Raguz^a ; ^c ; ¹ ; William J. O'Brien^b ; Witold K. Subczynski^a ; ^{subczyn@mcw.edu}
• 关键词：cholesterol ; membrane domains ; fluidity ; hydrophobic barrier ; oxygen permeation ; EPR ; spin-labeling
• 刊名：Experimental Eye Research
• 出版年：2012
• 期刊代码：293_00144835
• 类别：med
• 出版时间：April, 2012
• 卷：97
• 期：1
• 页码：117-129
• 文件大小：1742 K

摘要

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鈥攏amely, the domain formed by boundary lipids and the domain formed by trapped lipids鈥攚ere 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.