Andersen C, Jordy M, Benz R
Lehrstuhl fur Biotechnologie, Theodor-Boveri-Institut (Biozentrum), Universität Würzburg, Germany.
J Gen Physiol. 1995 Mar;105(3):385-401. doi: 10.1085/jgp.105.3.385.
LamB (maltoporin) of Escherichia coli outer membrane was reconstituted into artificial lipid bilayer membranes. The channel contains a binding site for sugars and is blocked for ions when the site is occupied by a sugar. The on and off reactions of sugar binding cause an increase of the noise of the current through the channel. The sugar-induced current noise of maltoporin was used for the evaluation of the sugar-binding kinetics for different sugars of the maltooligosaccharide series and for sucrose. The on rate constant for sugar binding was between 10(6) and 10(7) M-1.s-1 for the maltooligosaccharides and corresponds to the movement of the sugars from the aqueous phase to the central binding site. The off rate (corresponding to the release of the sugars from the channel) decreased with increasing number of glucose residues in the maltooligosaccharides from approximately 2,000 s-1 for maltotriose to 180 s-1 for maltoheptaose. The kinetics for sucrose movement was considerably slower. The activation energies of the stability constant and of the rate constants for sugar binding were evaluated from noise experiments at different temperatures. The role of LamB in the transport of maltooligosaccharides across the outer membrane is discussed.
将大肠杆菌外膜的LamB(麦芽糖孔蛋白)重组到人工脂质双分子层膜中。该通道含有一个糖结合位点,当该位点被糖占据时,离子通道被阻断。糖结合的开启和关闭反应会导致通过通道的电流噪声增加。麦芽糖孔蛋白的糖诱导电流噪声用于评估麦芽寡糖系列不同糖类以及蔗糖的糖结合动力学。麦芽寡糖的糖结合开启速率常数在10⁶至10⁷ M⁻¹·s⁻¹之间,这对应于糖从水相转移到中央结合位点的过程。麦芽寡糖中葡萄糖残基数量增加时,关闭速率(对应于糖从通道释放)从麦芽三糖的约2000 s⁻¹降至麦芽七糖的180 s⁻¹。蔗糖移动的动力学要慢得多。通过在不同温度下的噪声实验评估了糖结合的稳定常数和速率常数的活化能。讨论了LamB在麦芽寡糖跨外膜转运中的作用。