Carpi-Medina P, León V, Espidel J, Whittembury G
Venezuelan Institute of Scientific Investigation, IVIC, Caracas.
J Membr Biol. 1988 Aug;104(1):35-43. doi: 10.1007/BF01871900.
The diffusive water permeability (Pd) of the plasma membrane of proximal kidney tubule cells was measured using a 1H-NMR technique. The values obtained for the exchange time (Tex) across the membrane were independent of the cytocrit and of the Mn2+ concentration (in the range 2.5 to 5 mM). At 25 degrees C the calculated Pd value was (per cm2 of outer surface area without taking into account membrane invaginations) 197 +/- 17 microns/sec. This value equals 22.3 +/- 1.9 microns/sec when the invaginations are taken into account. Cell exposure to 2.5 mM parachloromercuribenzenesulfonic acid, pCMBS, (for 20 to 35 min) reduced Pd to 45% of its control value. Five mM dithiothreitol, DTT, reverted this effect. The activation energy for the diffusive water flux was 5.2 +/- 1.0 kcal/mol under control conditions. It increased to 9.1 +/- 2.2 kcal/mol in the presence of 2.5 mM pCMBS. Using our previous values for the osmotic water permeability (Pos) in proximal straight tubular cells the Pos/Pd ratio equals 18 +/- 1, under control conditions, and 3.2 +/- 0.3 in the presence of pCMBS. These experimental results indicate the presence of pathways for water, formed by proteins, crossing these membranes, which are closed by pCMBS. Assuming laminar flow (within the pore), from Pos/Pd of 13 to 18 an unreasonably large pore radius of 12 to 15 A is calculated which would not hinder cell entry of known extracellular markers. Alternatively, for a single-file pore, 11 to 20 would be the number of water molecules which would be in tandem inside the pore.(ABSTRACT TRUNCATED AT 250 WORDS)
利用1H-NMR技术测量了近端肾小管细胞质膜的扩散水渗透率(Pd)。跨膜交换时间(Tex)的测量值与血细胞比容和Mn2+浓度(2.5至5 mM范围内)无关。在25℃时,计算得到的Pd值(每平方厘米外表面面积,未考虑膜内陷)为197±17微米/秒。考虑膜内陷时,该值等于22.3±1.9微米/秒。细胞暴露于2.5 mM对氯汞苯磺酸盐(pCMBS)中(20至35分钟),Pd降低至对照值的45%。5 mM二硫苏糖醇(DTT)可逆转此效应。在对照条件下,扩散水通量的活化能为5.2±1.0千卡/摩尔。在2.5 mM pCMBS存在下,活化能增加至9.1±2.2千卡/摩尔。根据我们之前测得的近端直小管细胞的渗透水渗透率(Pos)值,对照条件下Pos/Pd比值为18±1,在pCMBS存在下为3.2±0.3。这些实验结果表明,存在由蛋白质形成的水通道穿过这些膜,这些通道可被pCMBS封闭。假设为层流(在孔内),根据Pos/Pd为13至18计算出的孔半径大得不合理,为12至15埃,这不会阻碍已知细胞外标志物进入细胞。或者,对于单排孔,孔内串联的水分子数为11至20个。(摘要截短于250字)