BOOKCHIN

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。主要的长期目标是彻底了解镰状细胞(SS)病的分子和细胞病理生理学。其中一个项目专注于HB S与红细胞膜相互作用改变细胞功能的机制，从而导致临床溶血性贫血和广泛的微血管闭塞。我们的近期目标是：1.研究镰刀样离子通透途径(S)的功能性质和可能的分子本质；2.研究我们发现的高Na+、低K+、低密度、阳离子渗漏SS和正常红细胞的产生机制(S)；我们的假设是，循环SS和变异红细胞之间的体积和密度(或细胞Hb浓度)的显著异质性是网织红细胞转运系统异质性的结果，并受到Hb膜相互作用的直接和间接影响。IV.研究各种转基因镰刀鼠模型的Hb聚合性和红细胞离子转运功能，以确定最适合检测镰刀细胞病的各种病理生理机制和治疗手法的模型。一旦确定了特征，这些将服务于上述三个目标的调查。在第二个项目中，我们将：V.继续表征脱氧-Hb S聚合物、纤维和凝胶的分子相互作用部位，重点是通过半合成方法或直接化学修饰对特定修饰敏感的潜在重要部位；关于聚合物不同结构水平上特定残基的顺式或反式作用；以及非S Hb参与或改变聚合过程的方式；Vi.进一步发展我们新的聚合物平衡溶解度的微观排除体积分析，右旋糖苷-Csat；vii.测试CP的恒定性或变异性，即Hb在聚合物中的浓度；检查2，3-DPG、ATP、RBC酶和小分子在PWC、Sol和膜中的分配情况；评估红细胞膜的正常或异常(镰状细胞)成分在细胞内聚合物的动力学和组装中的作用；以及X.通过直接观察红细胞膜中脱氧Hb S纤维的形成来研究。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The main long-term goal is a thorough understanding of the molecular and cellular pathophysiology of sickle cell (SS) disease. One project focuses on the mechanisms by which Hb S interacts with the RBC membranes to alter cell functions, contributing to clinical hemolytic anemia and widespread microvascular occlusion. Our immediate aims are: I. To characterize the functional properties and possible molecular nature of the sickling-induced ion permeability pathway(s), "Psickle";II. To investigate the mechanism(s) of generation of the high-Na+, low-K+, low-density, cation-leaky SS and normal RBCs we discovered; Ill. To pursue our hypothesis that the marked heterogeneity of volume and density (or cell Hb concentration) among circulating SS and variant RBCs results from heterogeneity of transport systems in the reticulocytes, with the direct and indirect effects of Hb membrane interactions. IV. To characterize the Hb-polymerization properties and RBC ion-transport; functions of a variety of transgenic sickle mouse models to identify those most suitable to test various pathophysiological mechanisms and therapeutic maneuvers in sickle cell disease. Once characterized, these will serve investigations within the above three aims. In the second project, we will: V. Continue to characterize the molecular interaction sites of the deoxy-Hb S polymers, fibers and gels, focusing on potentially important sites susceptible to specific modification, by semisynthetic approaches or direct chemical modification; on the cis or trans role of specific residues at different structural levels of the polymer; and on the way non-S Hbs participate in or alter the polymerization process; VI. develop further our new micro- excluded volume assay of the equilibrium solubility of the polymer, the dextran-Csat; VII. Test the constancy or variability of Cp, the Hb concentration in the polymer; VIII. Examine the partitioning in the PWC, sol and membrane of 2,3-DPG, ATP, RBC enzymes and small molecules; IX. Assess the role of normal or abnormal (sickle cell) components of the RBC membrane on the kinetics and assembly of intracellular polymer; and X. Study by direct observation the formation of deoxy-Hb S fibers in RBCs.