Laboratory of Cellular and Molecular Physiology, Department of Biotechnology and Life Sciences, University of Insubria, Via Dunant 3, 21100, Varese, Italy.
Pflugers Arch. 2012 Aug;464(2):183-91. doi: 10.1007/s00424-012-1125-8. Epub 2012 Jun 23.
The effects of temperature on the functional properties of the intestinal oligopeptide transporter PepT1 from rabbit have been investigated using electrophysiological methods. The dipeptide Gly-Gln at pH 6.5 or 7.5 was used as substrate. Raising the temperature in the range 20-30 °C causes an increase in the maximal transport-associated current (I (max)) with a Q (10) close to 4. Higher temperatures accelerate the rate of decline of the presteady-state currents observed in the absence of organic substrate. The voltage dependencies of the intramembrane charge movement and of the time constant of decline are both shifted towards more negative potentials by higher temperatures. The shift is due to a stronger action of temperature on the outward rate of charge movement compared to the inward rate, indicating a lower activation energy for the latter process. Consistently, the activation energy for the complete cycle is similar to that of the inward rate of charge movement. Temperature also affects the binding rate of the substrate: the K (0.5) -V curve is shifted to more negative potentials by higher temperatures, resulting in a lower apparent affinity in the physiological range of potentials. The overall efficiency of transport, estimated as the I (max)/K (0.5) ratio is significantly increased at body temperature.
采用电生理学方法研究了温度对兔肠寡肽转运蛋白 PepT1 功能特性的影响。在 pH6.5 或 7.5 时使用二肽 Gly-Gln 作为底物。在 20-30°C 的范围内升高温度会导致与最大转运相关的电流 (I(max)) 增加,Q(10)接近 4。较高的温度会加速在没有有机底物存在时观察到的稳态前电流的下降速率。较高的温度会使跨膜电荷移动的电压依赖性和下降时间常数向更负的电位移动。这种移位是由于温度对外向电荷移动速率的作用比对内向电荷移动速率的作用更强,表明后者过程的活化能较低。一致地,完整循环的活化能与内向电荷移动速率的活化能相似。温度还会影响底物的结合速率:较高的温度会使 K(0.5)-V 曲线向更负的电位移动,导致在生理电位范围内的表观亲和力降低。作为 I(max)/K(0.5) 比值的转运总效率在体温下显著增加。