TRANSMEMBRANOUS INCORPORATION OF PHOTOELECTRICALLY ACTIVE BACTERIORHODOPSIN IN PLANAR LIPID BILAYERS
TRANSMEMBRANOUS INCORPORATION OF PHOTOELECTRICALLY ACTIVE BACTERIORHODOPSIN IN PLANAR LIPID BILAYERS
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DOI：
10.1073/pnas.78.12.7502
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
HEYN, MP
中科院分区：
文献类型：
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作者：
BAMBERG, E;DENCHER, NA;HEYN, MP
报道了在黑脂膜中掺入嗜盐细菌紫质的各种方法。以紫色膜贴片和单体细菌视紫红质为起始材料。细菌紫质结合到平面脂质双分子层是通过以下方法实现的。紫色膜块从水中转移到正构烷烃中的脂质溶液中。黑色的膜由这种有机悬浮液形成。将含有溶剂和紫色膜的脂质层铺在空气/水界面上。这些层被用来形成平面双层。含有紫色膜或单体细菌视紫红质的囊泡在空气/水界面上扩散，并从产生的层形成双层。在光照下，三种情况下均观察到稳态光电流，表明这些方法导致蛋白质在平面黑色脂质膜上的功能性跨膜整合。研究了外加电场对方法1所形成膜的抽运过程的影响。200mv，光电流趋于零。通过在洗浴液中加入含有单体细菌视紫红质的囊泡，可以使平面脂质双分子层具有光电活性。在这种情况下，只观察到瞬时光电流，这表明囊泡附着在黑色脂质膜上，但没有与之融合。
Various methods to incorporate Halobacterium halobium bacteriorhodopsin in black lipid membranes are reported. Both purple membrane patches and monomeric bacteriorhodopsin were used as starting material. The incorporation of bacteriorhodopsin into planar lipid bilayers was achieved by the following methods. Purple membrane patches were transferred from water to solutions of lipids in n-alkanes. Black membranes were formed from such organic suspensions. Lipid layers containing solvent and purple membranes were spread on an air/water interface. These layers were used to form planar bilayers. Vesicles containing purple membranes or monomeric bacteriorhodopsin were spread on an air/water interface and, from the resulting layer, bilayers were formed. On illumination, steady-state photocurrents were observed in all 3 cases, indicating that these methods lead to functional transmembraneous integration of the protein in the planar black lipid membrane. The influence of an applied electric field on the pumping process was studied on membranes formed by using method i. At .apprx. 200 mV, the photocurrent tends to zero. It was possible to make planar lipid bilayers photoelectrically active by adding vesicles containing monomeric bacteriorhodopsin to the bathing solution. In this case, only transient photocurrents were observed, suggesting that the vesicles are attached to but not fused with the black lipid membrane.