Electrical manipulation of glycan phosphatidyl inositol tethered proteins in planar supported bilayers
Electrical manipulation of glycan phosphatidyl inositol tethered proteins in planar supported bilayers
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DOI：
10.1016/s0006-3495(96)79462-6
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发表时间：
1996-11-01
影响因子：
3.4
通讯作者：
Boxer, SG
中科院分区：
文献类型：
--
作者：
Groves, JT;Wulfing, C;Boxer, SG
电场已被用于操纵和浓缩糖甘-磷脂酰肌醇(GPI)-系固蛋白在平面支撑双层。研究了自然GPI连接的CD48，以及工程形式的I-E(k)和B7-2，其中它们的跨膜结构域已被GPI连接遗传地取代。用荧光标记的抗体标记蛋白质，使电场诱导的行为可以用荧光显微镜观察。这三种蛋白复合物都通过单个GPI连接体B7-2和CD48向阴极迁移，其移动速度明显快于具有两个GPI连接体的I-E(k)。刻划在膜上的图案作为横向扩散的屏障，并用于将蛋白质分离成高度集中的畜栏。所有场诱导的浓度曲线都是完全可逆的，这表明支持的双分子层提供了一个稳定的流体环境，gpi拴系蛋白可以在其中被操纵。电控制膜系蛋白空间分布的能力为生物膜的研究和膜基器件的发展提供了新的机会。
Electric fields have been used to manipulate and concentrate glycan-phosphatidyl inositol (GPI)-tethered proteins in planar supported bilayers. Naturally GPI-linked CD48, along with engineered forms of I-E(k) and B7-2, in which their transmembrane domains have been genetically replaced with the GPI linkage, were studied. The proteins were labeled with fluorescently tagged antibodies, allowing the electric field-induced behavior to be followed by epifluorescence microscopy. All three protein complexes were observed to migrate toward the cathode with B7-2 and CD48, each tethered to the membrane ky a single GPI linker, moving significantly faster the the I-E(k), which has two GPI linkers. Patterns scratched into the membrane function as barriers to lateral diffusion and were used to isolate the proteins into highly concentrated corrals. All field-induced concentration profiles were completely reversible, indicating that the supported bilayer provides a stable, fluid environment in which GPI-tethered proteins can be manipulated. The ability to electrically control the spatial distribution of membrane-tethered proteins provides new opportunities for the study of biological membranes and the development of membrane-based devices.