Hofrichter J, Ross P D, Eaton W A
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4864-8. doi: 10.1073/pnas.71.12.4864.
We report the results of a kinetic investigation on the gelation of purified deoxyhemoglobin S. Gelation was induced by raising the temperature and was monitored by measuring both the heat absorbed, with a microcalorimeter, and the appearance of linear birefringence, with a microspectrophotometer. The kinetics are unusual. Prior to the onset of gelation there is a delay period, followed by a sigmoidal progress curve. The delay time is formally dependent on approximately the 30th power of the deoxyhemoglobin S concentration; a decrease in concentration from 23 to 22 g/dl increases the delay time by a factor of four. It is also extremely temperature dependent; a 1 degrees C temperature rise in the range 20-30 degrees C almost halves the delay time. From these results we conclude that the initial rate is controlled by the nucleation of individual fibers. We present a kinetic model that accounts for the concentration, temperature, and time dependence of the initial phase of the gelation reaction. Extrapolation of our data to physiological conditions predicts that changes in intracellular hemoglobin concentration and oxygen saturation, realizable in vivo, produce enormous changes in the delay time. The range of delay times spans both the mean capillary transit and total circulation times. This result points to the delay time as an extremely important variable in determining the course of sickle cell disease, and suggests a new approach to therapy.
我们报告了对纯化的脱氧血红蛋白S凝胶化的动力学研究结果。通过升高温度诱导凝胶化,并使用微量量热计测量吸收的热量以及使用显微分光光度计测量线性双折射的出现来进行监测。动力学情况不同寻常。在凝胶化开始之前有一个延迟期,随后是一个S形的进展曲线。延迟时间形式上大约取决于脱氧血红蛋白S浓度的30次方;浓度从23克/分升降至22克/分升会使延迟时间增加四倍。它也极其依赖温度;在20 - 30摄氏度范围内温度升高1摄氏度几乎会使延迟时间减半。根据这些结果我们得出结论,初始速率由单个纤维的成核作用控制。我们提出了一个动力学模型,该模型解释了凝胶化反应初始阶段的浓度、温度和时间依赖性。将我们的数据外推到生理条件预测,体内可实现的细胞内血红蛋白浓度和氧饱和度的变化会使延迟时间产生巨大变化。延迟时间的范围涵盖了平均毛细血管运输时间和总循环时间。这一结果表明延迟时间是决定镰状细胞病病程的一个极其重要的变量,并提示了一种新的治疗方法。