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从大肠杆菌中分离的膜囊泡中6-磷酸葡萄糖的转运:外加电势和pH梯度的影响

Glucose 6-phosphate transport in membrane vesicles isolated from Escherichia coli: effect of imposed electrical potential and pH gradient.

作者信息

LeBlanc G, Rimon G, Kaback H R

出版信息

Biochemistry. 1980 May 27;19(11):2522-8. doi: 10.1021/bi00552a034.

Abstract

Imposition of a membrane potential (delta psi, interior negative) or a pH gradient (delta pH, interior alkaline) across the membrane of Escherichia coli DF2000 leads to a marked, transient increase in glucose 6-phosphate transport that varies systematically with pH. Outwardly directed potassium diffusion gradients in the presence of valinomycin (i.e., generation of delta psi, interior negative) drive glucose 6-phosphate transport at pH 7.5 but much less effectively at pH 5.5, although the magnitudes of the transient delta psi generated are comparable at both pH values. Similarly, imposition of delta psi (interior negative) retards the rate of passive, carrier-mediated glucose 6-phosphate efflux down a concentration gradient at pH 7.5 but not at pH 5.5. In contrast, imposition of delta pH (interior alkaline) by means of outwardly directed acetate diffusion gradients drives glucose 6-phosphate accumulation at pH 5.5 but is relatively ineffective at pH 7.5. The results are independent of the pK of glucose 6-phosphate and provide strong support for the argument that the glucose 6-phosphate porter catalyzes an electrically neutral reaction at acid pH and an electrogenic reaction at alkaline pH. In addition, they are entirely consistent with the hypothesis that the proton/glucose 6-phosphate stoichiometry increases at alkaline pH [Rottenberg, H. (1976) FEBS Lett. 66, 159; Ramos, S., & Kaback, H.R. (1977) Biochemistry 16, 854, 4271].

摘要

在大肠杆菌DF2000的细胞膜上施加膜电位(Δψ,胞内为负)或pH梯度(ΔpH,胞内为碱性)会导致6-磷酸葡萄糖转运显著且短暂地增加,这种增加随pH值呈系统性变化。在缬氨霉素存在的情况下,外向的钾扩散梯度(即产生Δψ,胞内为负)在pH 7.5时驱动6-磷酸葡萄糖的转运,但在pH 5.5时效果要差得多,尽管在这两个pH值下产生的瞬时Δψ大小相当。同样,施加Δψ(胞内为负)会在pH 7.5时减缓6-磷酸葡萄糖沿浓度梯度被动的、载体介导的外流速率,但在pH 5.5时则不会。相比之下,通过外向的乙酸盐扩散梯度施加ΔpH(胞内为碱性)在pH 5.5时驱动6-磷酸葡萄糖的积累,但在pH 7.5时相对无效。这些结果与6-磷酸葡萄糖的pK无关,并为以下观点提供了有力支持:6-磷酸葡萄糖转运体在酸性pH下催化电中性反应,在碱性pH下催化生电反应。此外,它们与质子/6-磷酸葡萄糖化学计量在碱性pH下增加的假设完全一致[罗滕贝格,H.(1976年)《欧洲生物化学学会联合会快报》66,159;拉莫斯,S.,& 卡巴克,H.R.(1977年)《生物化学》16,854,4271]。

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