SICKLE HEMOGLOBIN PATHOPHYSIOLOGY--STRUCTURES AND KINETICS

镰状血红蛋白病理生理学——结构和动力学

• 批准号: 6606072
• 负责人: FRANK A FERRONE
• 金额: $ 21.11万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6606072
• 关键词: cell morphology; chemical kinetics; cytoskeleton; erythrocyte membrane; gel; hemodynamics; hemoglobin Ss; ligands; oxygen tension; polymerization; protein isoforms; protein structure function; solubility; spectrin; thermodynamics

DESCRIPTION (provided by applicant): We propose to continue our studies on the mechanisms of polymerization and its connection to the origin of the pathophysiology of sickle cell disease. We are addressing five issues: 1) Participation of the red cell in nucleation of sickle cell hemoglobin. We have developed a unique capability that has very high precision of determining the effects of membranes on homogeneous nucleation, and will continue our studies on different types of membrane preparation. 2) Mixture Theory for Homogeneous Nucleation: Under-standing the kinetics of mixtures is critical, yet the theory of mixtures has not been expanded to include present models. 3) Identities and strengths of the energetically significant contacts in the polymer. The structure of the polymer is complex. We will measure solubilities for various mutants, as well as solubilities of mixtures of the mutants with HbS. In addition we will measure homogeneous and heterogeneous nucleation rates. From homogeneous nucleation rates and solubilities we can separate the contact energy from the vibrational entropy. Use of mixtures will allow us to ascertain cis-trans issues or inequivalence issues. 4) Tertiary effects on polymerization. This work will study the effect of ligands within the T quaternary structure, with particular emphasis on NOHbS. and 5) Dependence of gel rheology on location within a domain and relation to single fiber behavior. To correlate with single fiber studies, we will make measurements on polymer domains using thermally driven fluctuations. Experiments will use particle tracking methods to compare different regions of polymer domains. Mutants will be used to compare with predicted effects. We will observe the speed with which rigidification propagates through the domain.

描述（由申请人提供）： 我们建议继续研究聚合机制及其与 镰状细胞病病理生理学的起源。我们正在解决五个问题：1） 红细胞参与镰状细胞血红蛋白的成核。我们开发了一个 独特的能力，具有非常高的精度来确定膜的影响 均匀成核，并将继续我们对不同类型膜的研究 准备。 2) 均质成核的混合理论：理解动力学 混合物至关重要，但混合物理论尚未扩展到包括现有模型。 3) 聚合物中重要能量接触的特性和强度。结构 聚合物的结构很复杂。我们将测量各种突变体的溶解度以及溶解度 突变体与 HbS 的混合物。此外，我们将测量同质和 异质成核率。从均匀成核率和溶解度我们可以 将接触能与振动熵分开。使用混合物将使我们能够 确定顺反问题或不平等问题。 4) 对聚合的三级效应。这项工作将研究 T 四级结构内配体的影响，特别是 NOHbS。 5) 凝胶流变学对域内位置的依赖性以及与单个区域的关系 纤维行为。为了与单纤维研究相关联，我们将对聚合物进行测量 使用热驱动波动的域。实验将使用粒子跟踪方法 比较聚合物域的不同区域。突变体将被用来与 预测的效果。我们将观察刚性化通过 领域。