MECHANISM OF POLYMERIZATION OF HB S

这项研究的目的是更好地了解 用生化方法研究脱氧血红蛋白S的聚合机理， 生物物理学，和重组DNA技术，与远程 目标是找到抑制脱氧聚合的方法- HB S.试图进一步阐明其作用机制。 聚合脱氧-血红蛋白S，我们将继续研究其作用 血红蛋白S聚合前的晶核组成及表征 使用包括动力学在内的各种生物物理技术的原子核 最近获奖的激光散射式光谱仪 我们的实验室。我们将澄清以下概率因素 不同杂化血红蛋白在镰刀状混合物中的成核作用 和非镰状血红蛋白，这可能与临床有关 非镰状血红蛋白异常患者的严重程度。 最近，我们通过以下方法成功地表达了人β-珠蛋白 重组DNA工程技术。我们现在要准备 不同的血红蛋白突变体，在 β6位置以及在分子间接触点 脱氧-HB S聚合物。利用这些血红蛋白，我们将研究 不同氨基酸对动力学和热力学的影响 聚合反应和脱氧Hb S的溶解性 最近的研究表明，在某些实验条件下 高温合成脱氧血红蛋白S聚合产物的条件 磷酸盐缓冲液类似于在低磷条件下形成的缓冲液 缓冲区。使用高磷酸盐缓冲液是有利的，因为 只需要少量的血红蛋白就可以运动 实验。 为了测量蛋白质的表面疏水性，我们将使用 高效凝胶过滤柱，TSK-Gel-SW，用于转换 涉及高磷酸盐缓冲液中的疏水相互作用柱。我们 将研究血红蛋白的表面疏水性和 乳清蛋白表面疏水性与溶解度的关系 各种血红蛋白。我们还将澄清两国之间的关系 疏水相互作用的强度和水溶液的溶解度 脱氧-HbS。这些研究将增加我们对 镰状细胞病的病理学，并可能有助于 发展一种分子方法来改善这种生活- 威胁着遗传性疾病。

The objective of this research is to better understand the mechanism of polymerization of deoxy-Hb S using biochemical, biophysical, and recombinant DNA technology, with the long-range goal of finding the means to inhibit the polymerization of deoxy- Hb S. In an attempt to further clarify the mechanism of polymerization of deoxy-Hb S, we will continue to study the role of nuclei prior to polymerization of Hb S and to characterize nuclei using various biophysical techniques including a dynamic laser light-scattering phototmeter which was recently awarded to our laboratory. We will clarify the probability factor for nucleation of various hybrid hemoglobins in the mixtures of sickle and non-sickle hemoglobins, which may be related to clinical severity of patients having with abnormal non-sickle hemoglobins. Recently, we succeeded in the expression of human beta-globin by recombinant DNA engineering techniques. We will now prepare various hemoglobin mutants with amino acid substitutions at the beta 6 position as well as at the intermolecular contact points of deoxy-Hb S polymers. Using these hemoglobins, we will study the effect of various amino acids on the kinetics and thermodynamics of polymerization and on the solubility of deoxy-Hb S. Other recent studies have shown that under certain experimental conditions polymerized products of deoxy-Hb S prepared in high phosphate buffer were similar to those formed in low phosphate buffer. The use of high phosphate buffer is advantageous, since only small amounts of hemoglobin are required for kinetic experiments. To measure the surface hydrophobicity of proteins, we will use a high performance gel filtration column, TSK-GEL-SW, which converts to a hydrophobic interaction column in high phosphate buffer. We will study the surface hydrophobicity of hemoglobins and the relationship between the surface hydrophobicity and solubility of various hemoglobins. We will also clarify the relationship between the strength of hydrophobic interaction and the solubility of deoxy-Hb S. These studies will add to our knowledge of the pathology of sickle cell disease and may contribute to the development of a molecular approach to ameliorating this life- threatening genetic disorder.