Biological membrane modeling with a liquid/liquid interface. Probing mobility and environment with total internal reflection excited fluorescence.
Biological membrane modeling with a liquid/liquid interface. Probing mobility and environment with total internal reflection excited fluorescence.
复制标题
具有液/液界面的生物膜建模。
DOI：
10.1016/s0006-3495(87)83225-3
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发表时间：
1987
影响因子：
3.4
通讯作者：
Gregorio Weber
中科院分区：
文献类型：
--
作者：
Larry E. Morrison;Gregorio Weber
激发光的全内反射，结合荧光强度和偏振测量，被用来选择性地研究荧光化合物吸附到两种不混溶液体之间的界面区域。构建了一个荧光计，它提供了在可变的入射角的激发，并允许从界面发射的偏振荧光的灵敏检测。研究了化合物4，4 ′-双-1-苯胺基-8-萘磺酸盐（bis-ANS）在十氢萘/水界面上的吸附行为，发现其对界面区具有显著的亲合力，大部分吸附分子位于十氢萘相中。吸附的荧光团显示出一个明显的受阻旋转的界面与旋转扩散系数3- 12倍低于预期的双ANS在溶液中的平面。虽然没有发现其他染料检查是显着的表面活性，阳离子表面活性剂的加入足以诱导阴离子荧光团1-氨基萘-3，6，8-三磺酸的吸附。发现这种荧光孔在结合到界面时驻留在水性环境中，并且它还表现出在界面平面中的受阻旋转。随着染料浓度的增加，两种吸附染料均表现出与激发能量转移一致的极化降低。通过加入牛血清白蛋白，双ANS的吸附被逆转。细胞色素b5的膜蛋白被发现不结合在十氢萘/水界面，表明与脂质的相互作用是必需的，其粘附到生物膜。
Total internal reflection of exciting light, in combination with fluorescence intensity and polarization measurements, was used to selectively study fluorescent compounds adsorbed to the interface region between two immiscible liquids. A fluorometer was constructed which provided excitation at variable angles of incidence and allowed sensitive detection of polarized fluorescence emitted from the interface. The compound 4,4'-bis-1-phenylamino-8-naphthalenesulfonate (bis-ANS) was examined at a decalin/water interface and was found to possess remarkable affinity for the interface region with the bulk of the adsorbed molecule residing in the decalin phase. The adsorbed fluorophore displayed an apparent hindered rotation in the plane of the interface with a rotational diffusion coefficient 3- to 12-fold lower than that expected for bis-ANS in solution. While other dyes examined were not found to be significantly surface active, the addition of cationic surfactant sufficed to induce adsorption of the anionic fluorophore 1-aminonaphthalene-3,6,8-trisulfonic acid. This fluoropore was found to reside in an aqueous environment when bound to the interface, and it also exhibited hindered rotation in the plane of the interface. As the concentrations of the dyes were increased, both adsorbed dyes exhibited polarization reductions consistent with excitation energy transfer. Adsorption of bis-ANS was reversed by addition of bovine serum albumin. The membrane protein cytochrome b5 was found not to bind at the decalin/water interface, indicating that interaction with lipid is required for its adherence to biological membranes.