Benz R, Schmid A, Vos-Scheperkeuter G H
Lehrstuhl für Biotechnologie der Universität Würzburg, Federal Republic of Germany.
J Membr Biol. 1987;100(1):21-9. doi: 10.1007/BF02209137.
Lipid bilayer experiments were performed with the sugar-specific LamB (maltoporin) channel of Escherichia coli outer membrane. Single-channel analysis of the conductance steps caused by LamB showed that there was a linear relationship between the salt concentration in the aqueous phase and the channel conductance, indicating only small or no binding between the ions and the channel interior. The total or the partial blockage of the ion movement through the LamB channel was not dependent on the ion concentration in the aqueous phase. Both results allowed the investigation of the sugar binding in more detail, and the stability constants of the binding of a large variety of sugars to the binding site inside the channel were calculated from titration experiments of the membrane conductance with the sugars. The channel was highly cation selective, both in the presence and absence of sugars, which may be explained by the existence of carbonyl groups inside the channel. These carbonyl groups may also be involved in the sugar binding via hydrogen bonds. The kinetics of the sugar transport through the LamB channel were estimated relative to maltose by assuming a simple one-site, two-barrier model from the relative rates of permeation taken from M. Luckey and H. Nikaido (Proc. Natl. Acad. Sci. USA 77:165-171 (1980a)) and the stability constants for the sugar binding given in this study.
利用大肠杆菌外膜中对糖具有特异性的LamB(麦芽糖孔蛋白)通道进行了脂质双层实验。对LamB引起的电导步骤进行单通道分析表明,水相中的盐浓度与通道电导之间存在线性关系,这表明离子与通道内部之间只有少量结合或没有结合。离子通过LamB通道的全部或部分阻断并不取决于水相中的离子浓度。这两个结果使得能够更详细地研究糖结合情况,并通过用糖对膜电导进行滴定实验计算出多种糖与通道内结合位点结合的稳定常数。无论有无糖存在,该通道都具有高度的阳离子选择性,这可能是由于通道内部存在羰基所致。这些羰基也可能通过氢键参与糖结合。通过假设一个简单的单位点、双屏障模型,根据M. Luckey和H. Nikaido(《美国国家科学院院刊》77:165 - 171 (1980a))给出的相对渗透速率以及本研究中给出的糖结合稳定常数,相对于麦芽糖估算了糖通过LamB通道的转运动力学。