Spray D C, Saez J C, Brosius D, Bennett M V, Hertzberg E L
Proc Natl Acad Sci U S A. 1986 Aug;83(15):5494-7. doi: 10.1073/pnas.83.15.5494.
We have shown previously that conductance of rat liver gap junctions is blocked by an affinity-purified polyclonal antibody generated against rat liver junctional membranes, is not affected by moderate transjunctional or transmembrane potentials, and is reversibly decreased by cytoplasmic acidification and perfusion with octanol. We have now recorded currents from isolated liver gap junctions using patch electrodes dipped through a layer of mixed lipids whose concentrations match those of isolated liver appositional membranes. These currents are blocked by the same polyclonal antibody, are insensitive to moderate voltages imposed across the pipette tip, and are reversibly blocked by similar concentrations of H ions and octanol as are junctions in situ. The currents are likely to be gap junctional in origin; their block by low pH and other agents indicates that the gating mechanisms are intrinsic to the gap junctions themselves and presumably result from conformational change in the channel-forming protein.
我们之前已经表明,大鼠肝脏间隙连接的电导被一种针对大鼠肝脏连接膜产生的亲和纯化多克隆抗体所阻断,不受适度的跨连接或跨膜电位影响,并且通过细胞质酸化和用辛醇灌注而可逆地降低。我们现在使用浸过一层混合脂质的膜片电极记录了分离的肝脏间隙连接的电流,这些脂质的浓度与分离的肝脏并列膜的浓度相匹配。这些电流被相同的多克隆抗体阻断,对施加在移液管尖端的适度电压不敏感,并且被与原位连接类似浓度的氢离子和辛醇可逆地阻断。这些电流很可能起源于间隙连接;它们被低pH值和其他试剂阻断表明门控机制是间隙连接本身所固有的,大概是由形成通道的蛋白质的构象变化引起的。