Thilo L, Träuble H, Overath P
Biochemistry. 1977 Apr 5;16(7):1283-90. doi: 10.1021/bi00626a007.
In an attempt to understand the mechanism by which a structural change of membrane lipids affects transport functions, the temperature dependence of transport rate has been measured to below the low temperature end of the fluid in equilibrium ordered phase transition of the membrane lipids. The unsaturated fatty acid requiring Escherichia coli strain T105 was supplemented with either trans-delta9-octadecenoate or trans-delta9-hexadecenoate or supplemented with and subsequently starved for cis-delta9-octadecenoate. Fluid in equilibrium ordered phase transitions measured in whole cells using the fluorescence probe N-phenyl-1-naphthylamine were compared with the temperature dependence of beta-glucoside and beta-galactoside transport. In addition to the previously observed downward "break" in the Arrhenius plot of transport rate which occurred near the middle of the phase transition temperature range, a second upward "break" was observed which could be correlated with the low-temperature end of the phase transition. These experiments are interpreted in terms of a partitioning of transport proteins between ordered and fluid domains which is described by a lateral distribution coefficient, k. This distribution coefficient varies with the membrane lipid composition as well as with the transport system. Values for k suggest a 2-20-fold preference for the partitioning of transport proteins into the fluid parts of the membrane.
为了理解膜脂结构变化影响转运功能的机制,已测量了转运速率的温度依赖性,直至膜脂平衡有序相转变的低温端以下。对需要不饱和脂肪酸的大肠杆菌菌株T105补充反式-Δ9-十八碳烯酸或反式-Δ9-十六碳烯酸,或补充顺式-Δ9-十八碳烯酸后再使其饥饿。使用荧光探针N-苯基-1-萘胺在全细胞中测量的平衡有序相转变与β-葡萄糖苷和β-半乳糖苷转运的温度依赖性进行了比较。除了先前在相转变温度范围中间附近观察到的转运速率阿仑尼乌斯图中的向下“断点”外,还观察到第二个向上“断点”,它可能与相转变的低温端相关。这些实验根据转运蛋白在有序和流体区域之间的分配来解释,这种分配由横向分布系数k描述。该分布系数随膜脂组成以及转运系统而变化。k值表明转运蛋白分配到膜的流体部分的偏好性为2至20倍。