de Godoy C M, Cukierman S
Department of Physiology, Loyola University Medical Center, Maywood, Illinois 60153, USA.
Biophys J. 2001 Sep;81(3):1430-8. doi: 10.1016/s0006-3495(01)75798-0.
Proton conductance (g(H)) in single SS stereoisomers of dioxolane-linked gramicidin A (gA) channels were measured in different phospholipid bilayers at different HCl concentrations. In particular, measurements were obtained in bilayers made of 1,2-diphytanoyl 3-phosphocholine (DiPhPC) or its ethylated derivative 1,2-diphytanoyl 3-ethyl-phosphocholine (et-DiPhPC,). The difference between these phospholipids is that in et-DiPhPC one of the phosphate oxygens is covalently linked to an ethyl group and cannot be protonated. In relatively dilute acid solutions, g(H) in DiPhPC is significantly higher than in et-DiPhPC. At high acid concentrations, g(H) is the same in both diphytanoyl bilayers. Such differences in g(H) can be accounted for by surface charge effects at the membrane/solution interfaces. In the linear portion of the log g(H)-log [H] relationship, g(H) values in diphytanoyl bilayers were significantly larger (approximately 10-fold) than in neutral glyceryl monooleate (GMO) membranes. The slopes of the linear log-log relationships between g(H) and [H] in diphytanoyl and GMO bilayers are essentially the same (approximately 0.76). This slope is significantly lower than the slope of the log-log plot of proton conductivity versus proton concentration in aqueous solutions (approximately 1.00). Because the chemical composition of the membrane-channel/solution interface is strikingly different in GMO and diphytanoyl bilayers, the reduced slope in g(H)-[HCl] relationships may be a characteristic of proton transfer in the water wire inside the SS channel. Values of g(H) in diphytanoyl bilayers were also significantly larger than in membranes made of the more common biological phospholipids 1-palmitoyl 2-oleoyl phosphocholine (POPC) or 1-palmitoyl 2-oleoyl phosphoethanolamine (POPE). These differences, however, cannot be accounted for by different surface charge effects or by different internal dipole potentials. On the other hand, maximum g(H) measured in the SS channel does not depend on the composition of the bilayer and is determined essentially by the reduced mobility of protons in concentrated acid solutions. Finally, no experimental evidence was found in support of a lateral proton movement at the phospholipid/solution interface contributing to g(H) in single SS channels. Protein-lipid interactions are likely to modulate g(H) in the SS channel.
在不同盐酸浓度下,于不同磷脂双层中测量了二氧戊环连接的短杆菌肽A(gA)通道单一SS立体异构体的质子电导率(g(H))。具体而言,测量是在由1,2-二植烷酰基-3-磷酸胆碱(DiPhPC)或其乙基化衍生物1,2-二植烷酰基-3-乙基磷酸胆碱(et-DiPhPC)制成的双层膜中进行的。这些磷脂之间的差异在于,在et-DiPhPC中,一个磷酸氧原子与一个乙基共价连接,不能被质子化。在相对稀的酸性溶液中,DiPhPC中的g(H)显著高于et-DiPhPC中的g(H)。在高酸浓度下,两种二植烷酰基双层膜中的g(H)相同。g(H)的这种差异可由膜/溶液界面处的表面电荷效应来解释。在log g(H)-log [H]关系的线性部分,二植烷酰基双层膜中的g(H)值比中性单油酸甘油酯(GMO)膜中的g(H)值大得多(约10倍)。二植烷酰基双层膜和GMO双层膜中g(H)与[H]之间的线性对数-对数关系的斜率基本相同(约0.76)。该斜率显著低于水溶液中质子电导率与质子浓度的对数-对数图的斜率(约1.00)。由于GMO双层膜和二植烷酰基双层膜中膜-通道/溶液界面的化学组成显著不同,g(H)-[HCl]关系中斜率的降低可能是SS通道内水线中质子转移的一个特征。二植烷酰基双层膜中的g(H)值也显著大于由更常见的生物磷脂1-棕榈酰基-2-油酰基磷酸胆碱(POPC)或1-棕榈酰基-2-油酰基磷酸乙醇胺(POPE)制成的膜中的g(H)值。然而,这些差异不能用不同的表面电荷效应或不同的内部偶极势来解释。另一方面,在SS通道中测得的最大g(H)不依赖于双层膜的组成,基本上由浓酸溶液中质子迁移率的降低所决定。最后,没有发现实验证据支持磷脂/溶液界面处的横向质子运动对单一SS通道中的g(H)有贡献。蛋白质-脂质相互作用可能会调节SS通道中的g(H)。