Adachi K, Asakura T
Blood Cells. 1982;8(2):213-24.
Diluted solutions of deoxyhemoglobin S in concentrated phosphate buffer form aggregates or gels with a clear exhibition of a delay time. The aggregates can be liquified by cooling, bubbling with O2 or CO gas, or the dilution of phosphate buffer with water. These properties can be used as a simple method for studying the mechanism of polymerization and depolymerization of hemoglobins. The advantages of this method are: 1) The amount of hemoglobin sample required is only 1% to 5% of that required for the gelation of deoxy-Hb S in low phosphate buffer. 2) The kinetics can be measured turbidimetrically using an ordinary spectrophotometer. 3) The solubility of hemoglobin can be directly determined by taking the absorption spectrum of the supernatant solution after polymerization. 4) The polymer phase can be easily separated from the solution so that the amount and composition of the polymers can be analyzed. 5) The volume of the polymer phase is so small that excluded volume effect can be neglected. 6) The method can be applied to the study of polymerization of non-sickle hemoglobins and that of mixtures of sickle and non-sickle hemoglobins. The major question is whether the polymerization of hemoglobin in concentrated phosphate buffer is the same as that of deoxy-Hb S in low phosphate buffer. To answer this question, we studied the polymerization of Hb S, Hb A, Hb C Harlem, and Hb C in phosphate buffers of different molarities. We also studied the mechanism of the conversion of gels of these hemoglobins into crystals.
脱氧血红蛋白S在浓磷酸盐缓冲液中的稀释溶液会形成聚集体或凝胶，并明显表现出延迟时间。通过冷却、用氧气或一氧化碳气体鼓泡，或用水稀释磷酸盐缓冲液，聚集体可以液化。这些特性可作为研究血红蛋白聚合和解聚机制的一种简单方法。该方法的优点有：1）所需血红蛋白样品量仅为低磷酸盐缓冲液中脱氧血红蛋白S凝胶化所需量的1%至5%。2）动力学可以用普通分光光度计通过比浊法测量。3）通过测定聚合后上清液的吸收光谱，可以直接确定血红蛋白的溶解度。4）聚合物相可以很容易地从溶液中分离出来，从而可以分析聚合物的量和组成。5）聚合物相的体积非常小，以至于可以忽略排除体积效应。6）该方法可应用于非镰状血红蛋白的聚合以及镰状和非镰状血红蛋白混合物的聚合研究。主要问题是浓磷酸盐缓冲液中血红蛋白的聚合是否与低磷酸盐缓冲液中脱氧血红蛋白S的聚合相同。为了回答这个问题，我们研究了不同摩尔浓度磷酸盐缓冲液中血红蛋白S、血红蛋白A、哈莱姆血红蛋白C和血红蛋白C的聚合。我们还研究了这些血红蛋白凝胶转化为晶体的机制。
