Ter Kuile B H, Wiemer E A, Michels P A, Opperdoes F R
Research Unit for Tropical Diseases, International Institute of Cellular and Molecular Pathology, Brussels, Belgium.
Mol Biochem Parasitol. 1992 Oct;55(1-2):21-7. doi: 10.1016/0166-6851(92)90123-2.
The membrane potential and pH gradient over the plasma membrane of the protozoan parasite Trypanosoma brucei were measured with radioactive indicators in combination with the silicone oil centrifugation technique over a range of temperatures. At 37 degrees C a small membrane potential and pH gradient of similar magnitude, but of opposite polarity, were measured. The resulting electrochemical proton gradient was almost zero. However, when the temperature was lowered from 37 degrees C to 22 degrees C, the internal pH was kept constant independent of the external pH and a membrane potential of between -100 and -150 mV was measured, depending on the external pH. Measurements at various temperatures between 15 degrees C and 37 degrees C revealed that above 26 degrees C the membrane potential collapsed and that this collapse correlated with a sudden increase in membrane fluidity. The uptake of 2-deoxy-D-glucose and of pyruvate, which are both mediated by facilitated diffusion carriers in the plasma membrane of the trypanosome, were also affected by this sudden increase in fluidity of the membrane. The overall rate of the conversion of glucose into its metabolites, which is independent of the plasma membrane, varied only gradually. We conclude (i) that major changes occur in the plasma membrane of T. brucei around 26 degrees C, that affect all membrane related processes; (ii) that the electrochemical proton gradient plays a minor role in the energy metabolism of T. brucei when it resides in the bloodstream of the mammalian host at 37 degrees C; and (iii) that below 26 degrees C an electrochemical proton gradient is maintained over the plasma membrane.
利用放射性指示剂结合硅油离心技术,在一系列温度范围内测量了原生动物寄生虫布氏锥虫质膜上的膜电位和pH梯度。在37℃时,测量到一个幅度相似但极性相反的小膜电位和pH梯度。由此产生的电化学质子梯度几乎为零。然而,当温度从37℃降至22℃时,内部pH保持恒定,与外部pH无关,并且根据外部pH测量到的膜电位在-100至-150 mV之间。在15℃至37℃之间的不同温度下进行的测量表明,高于26℃时膜电位崩溃,并且这种崩溃与膜流动性的突然增加相关。锥虫质膜中由易化扩散载体介导的2-脱氧-D-葡萄糖和丙酮酸的摄取也受到膜流动性突然增加的影响。葡萄糖转化为其代谢产物的总体速率与质膜无关,仅逐渐变化。我们得出以下结论:(i)布氏锥虫的质膜在26℃左右发生重大变化,影响所有与膜相关的过程;(ii)当布氏锥虫处于哺乳动物宿主血液中37℃时,电化学质子梯度在其能量代谢中起次要作用;(iii)低于26℃时,质膜上维持电化学质子梯度。