Kanemasa T, Banba K, Kasai M
J Biochem. 1987 Apr;101(4):1025-32. doi: 10.1093/oxfordjournals.jbchem.a121944.
Voltage-gated anion channels in vesicles prepared from the electric organ of Narke japonica were studied using two methods. Ionic permeability was measured by the light scattering method, which could be used to measure the ion permeation of whole vesicles but only at a time scale of slower than about 0.1 s. The single channel conductances and permeability ratios for various ions were measured after fusing the vesicles to phospholipid bilayers. Both sets of results coincided, indicating that the anion channels observed with the planar bilayer method are the major route for anion passage in these vesicles. The channels showed anion selectivity and did not allow the permeation of cations such as K+ and choline+. The single channel conductance was 18 pS in 0.1 M Cl-. SCN- inhibited the conductance in a voltage-dependent reversible manner on both sides of a channel. SCN- may bind to the Cl- binding site in a channel and thus block it. 4,4'-Diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) blocked a channel on the cis (extracellular) side irreversibly. The number of anion channels per vesicle was estimated to be about 50. It was also shown that all anion channels in the vesicles were open at the very instance of fusion with planar membranes.
采用两种方法对从日本电鳐电器官制备的囊泡中的电压门控阴离子通道进行了研究。通过光散射法测量离子通透性,该方法可用于测量整个囊泡的离子渗透,但仅在慢于约0.1秒的时间尺度上。将囊泡与磷脂双层融合后,测量了各种离子的单通道电导和渗透率比。两组结果一致,表明用平面双层法观察到的阴离子通道是这些囊泡中阴离子通过的主要途径。这些通道表现出阴离子选择性,不允许K+和胆碱+等阳离子渗透。在0.1 M Cl-中,单通道电导为18 pS。SCN-以电压依赖性可逆方式在通道两侧抑制电导。SCN-可能与通道中的Cl-结合位点结合并因此阻断它。4,4'-二异硫氰基芪-2,2'-二磺酸(DIDS)不可逆地阻断顺式(细胞外)侧的通道。每个囊泡中阴离子通道的数量估计约为50个。还表明,囊泡中的所有阴离子通道在与平面膜融合的瞬间都是开放的。