Sharp R R, Sen R
Biophys J. 1982 Oct;40(1):17-25. doi: 10.1016/S0006-3495(82)84453-6.
NMR spin-lattice relaxation rates of solvent protons have been used to measure the water permeability coefficient of the chromaffin granule membrane. The technique involves labeling the chromaffin granule interior with Mn+2, which provides an efficient relaxation pathway for intravesicular solvent protons. Added Mn+2 spontaneously accumulates in the chromaffin granule matrix in the presence of the divalent cation-specific ionophore A23187 and is maintained against a large concentration gradient. In this way, the internal proton relaxation rate is readily augmented to values some 10(2)-10(3) times greater than that in the extravesicular water space. Transmembranal water transport permits solvent protons in the extravesicular water space, in which most of the observed NMR signal orginates, to sample the highly relaxive environment of the chromaffin granule matrix. By this process, water permeation shortens the observed relaxation rate. The diffusive water permeability coefficient of the chromaffin granule membrane has been measured over the temperature range 0-38 degrees C. The permeability coefficient measured at 25 degrees C is comparable to a previously reported value for planar lipid bilayers composed of ox brain lipids and cholesterol (Pd approximately equal to 0.37-0.53 10(-3)) cm X s-1 at 25 degrees C) but is substantially less than values for the plasma membranes of erythrocytes and Chlorella. Hypothesized hydrophilic "pores," thought to provide parallel permeation pathways in the latter membranes, appear to be absent in chromaffin granule membranes. The water permeation rate exhibits Arrhenius temperature behavior and does not reflect a phase transition at 32 degrees-34 degrees C observed previously in ESR spin-label studies of chromaffin granule ghosts.
已利用溶剂质子的核磁共振自旋晶格弛豫速率来测量嗜铬粒蛋白颗粒膜的水渗透系数。该技术包括用Mn²⁺标记嗜铬粒蛋白颗粒内部,这为囊泡内溶剂质子提供了一条有效的弛豫途径。在二价阳离子特异性离子载体A23187存在的情况下,添加的Mn²⁺会自发地在嗜铬粒蛋白颗粒基质中积累,并逆着较大的浓度梯度得以维持。通过这种方式,内部质子弛豫速率很容易增加到比囊泡外水空间中的值大10² - 10³倍左右。跨膜水运输使囊泡外水空间中的溶剂质子(大部分观察到的核磁共振信号起源于此)能够进入嗜铬粒蛋白颗粒基质的高弛豫环境。通过这个过程,水渗透缩短了观察到的弛豫速率。已在0 - 38℃的温度范围内测量了嗜铬粒蛋白颗粒膜的扩散水渗透系数。在25℃下测得的渗透系数与先前报道的由牛脑脂质和胆固醇组成的平面脂质双层的值相当(25℃时Pd约等于(0.37 - 0.53)×10⁻³ cm·s⁻¹),但远低于红细胞和小球藻质膜的值。推测在后者膜中提供平行渗透途径的亲水性“孔”在嗜铬粒蛋白颗粒膜中似乎并不存在。水渗透速率表现出阿累尼乌斯温度行为,并且不反映先前在嗜铬粒蛋白颗粒空壳的电子自旋共振自旋标记研究中观察到的32℃ - 34℃的相变。