Ives H E, Verkman A S
Am J Physiol. 1985 Dec;249(6 Pt 2):F933-40. doi: 10.1152/ajprenal.1985.249.6.F933.
H+ permeability (PH) of brush border membrane vesicles isolated from rabbit renal cortex was measured from the rate of collapse of preformed pH gradients using acridine orange fluorescence quenching. n-Alkanols increased PH from 0.005 to 0.1 cm/s in a dose-dependent manner. At 25 degrees C, PH increased to 0.01 cm/s at [n-alkanol] = 90 mM (butanol), 30 mM (pentanol), 7 mM (hexanol), and 1.8 mM (heptanol). Activation energy (Ea) of PH was 21.6 kcal/mol (5-50 degrees C), which decreased to 18.5 kcal/mol in the presence of either 200 mM butanol or 12 mM hexanol. Membrane fluidity was estimated from diphenylhexatriene anisotropy (r). n-Alkanols decreased r from 0.25 to 0.18 in a dose-dependent manner. At 25 degrees C, r = 0.22 at [n-alkanol] = 200 mM (butanol), 27 mM (pentanol), 9.5 mM (hexanol), and 2 mM (heptanol). The effects of n-alkanols on PH and r correlated well with known n-alkanol lipid-water partition coefficients. Similar increases in PH and decreases in r were observed for nonalkanol lipid anesthetics. The effects of n-alkanols on the Na+-H+ antiporter and on osmotically driven water transport were also studied. At concentrations of n-alkanol that resulted in a 10-fold increase in PH, there was no significant effect on either Na+-H+ exchange or water transport. These results suggest a lipid pathway for brush border H+ diffusion that is distinct from both the Na+-H+ antiporter and the water transport pathway.
采用吖啶橙荧光猝灭法,根据预先形成的pH梯度的崩塌速率,测定了从兔肾皮质分离的刷状缘膜囊泡的H⁺通透性(PH)。正构烷醇以剂量依赖方式使PH从0.005增加至0.1 cm/s。在25℃时,当[正构烷醇] = 90 mM(丁醇)、30 mM(戊醇)、7 mM(己醇)和1.8 mM(庚醇)时,PH增加至0.01 cm/s。PH的活化能(Ea)为21.6 kcal/mol(5 - 50℃),在存在200 mM丁醇或12 mM己醇时降至18.5 kcal/mol。通过二苯基己三烯各向异性(r)估计膜流动性。正构烷醇以剂量依赖方式使r从0.25降至0.18。在25℃时,当[正构烷醇] = 200 mM(丁醇)、27 mM(戊醇)、9.5 mM(己醇)和2 mM(庚醇)时,r = 0.22。正构烷醇对PH和r的影响与已知的正构烷醇脂质 - 水分配系数密切相关。对于非烷醇脂质麻醉剂,观察到类似的PH增加和r降低。还研究了正构烷醇对Na⁺ - H⁺反向转运体和渗透驱动的水转运的影响。在导致PH增加10倍的正构烷醇浓度下,对Na⁺ - H⁺交换或水转运均无显著影响。这些结果表明,刷状缘H⁺扩散存在一条脂质途径,该途径与Na⁺ - H⁺反向转运体和水转运途径均不同。