Zipp A, James T L, Kuntz I D, Shohet S B
Biochim Biophys Acta. 1976 Apr 23;428(2):291-303. doi: 10.1016/0304-4165(76)90037-4.
The temperature and cell volume dependence of the NMR water proton line-width, spin-lattice, and spin-spin relaxation times have been studied for normal and sickle erythrocytes as well as hemoglobin A and hemoglobin S solutions. Upon deoxygenation, the spin-spin relaxation time (T2) decreases by a factor of 2 for sickle cells and hemoglobin S solutions but remains relatively constant for normal cells and hemoglobin A solutions. The spin-lattice relaxation time (T1) shows no significant change upon deoxygenation for normal or sickle packed red cells. Studies of the change in the NMR linewidth, T1 and T2 as the cell hydration is changed indicate that these parameters are affected only slightly by a 10-20% cell dehydration. This result suggests that the reported 10% cell dehydration observed with sickling is not important in the altered NMR properties. Low temperature studies of the linewidth and T1 for oxy and deoxy hemoglobin A and hemoglobin S solutions suggest that the "bound" water possesses similar properties for all four species. The low temperature linewidth ranges from about 250 Hz at -15 degrees C to 500 Hz at -36 degrees C and analysis of the NMR curves yield hydration values near 0.4 g water/g hemoglobin for all four species. The low temperature T1 data go through a minimum at -35 degrees C for measurements at 44.4 MHz and -50 degrees C for measurements at 17.1 MHz and are similar for oxy and deoxy hemoglobin A and hemoglobin S. These similarities in the low temperature NMR data for oxy and deoxy hemoglobin A and hemoglobin S suggest a hydrophobically driven sickling mechanism. The room temperature and low temperature relaxation time data for normal and sickle cells are interpreted in terms of a three-state model for intracellular water. In the context of this model the relaxation time data imply that type III, or irrotationally bound water, is altered during the sickling process.
已经对正常红细胞和镰状红细胞以及血红蛋白A和血红蛋白S溶液的核磁共振水质子线宽、自旋晶格和自旋-自旋弛豫时间的温度和细胞体积依赖性进行了研究。脱氧后,镰状细胞和血红蛋白S溶液的自旋-自旋弛豫时间(T2)降低了2倍,但正常细胞和血红蛋白A溶液的该时间保持相对恒定。对于正常或镰状压积红细胞,脱氧后自旋晶格弛豫时间(T1)没有显著变化。随着细胞水合作用的改变对核磁共振线宽、T1和T2变化的研究表明,细胞脱水10%-20%对这些参数的影响很小。该结果表明,镰状化时报道的10%细胞脱水在核磁共振特性改变中并不重要。对氧合和脱氧血红蛋白A和血红蛋白S溶液的线宽和T1进行低温研究表明,“结合”水对所有四种物质具有相似的性质。低温线宽在-15℃时约为250Hz,在-36℃时为500Hz,对核磁共振曲线的分析得出所有四种物质的水合值接近0.4g水/g血红蛋白。低温T1数据在44.4MHz测量时在-35℃出现最小值,在17.1MHz测量时在-50℃出现最小值,氧合和脱氧血红蛋白A和血红蛋白S的数据相似。氧合和脱氧血红蛋白A和血红蛋白S的低温核磁共振数据中的这些相似性表明存在疏水驱动的镰状化机制。根据细胞内水的三态模型解释了正常细胞和镰状细胞的室温及低温弛豫时间数据。在该模型的背景下,弛豫时间数据表明,III型或非旋转结合水在镰状化过程中发生了改变。