STRUCTURE OF THE CDB3:ANKYRINR COMPLEX IN ERYTHROCYTES BY EPR

通过 EPR 分析红细胞中 CDB3: 锚蛋白复合物的结构

• 批准号: 8364096
• 负责人: ALBERT H BETH
• 金额: $ 0.08万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364096
• 关键词: Adaptor Signaling Protein; Amino Acids; Anion Exchangers (Proteins); Architecture; Blood Circulation; Blood capillaries; Cells; Complex; Cytoplasmic Tail; Data; Erythrocyte Membrane; Erythrocytes; Funding; Grant; Hemolytic Anemia; Human; Lead; Measurement; Membrane; National Center for Research Resources; Peripheral; Physiologic pulse; Principal Investigator; Proteins; Research; Research Infrastructure; Resolution; Resources; Shapes; Source; Spectrum Analysis; Structure; Techniques; Technology; United States National Institutes of Health; capillary; cost; protein complex; quantum; shear stress

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. A remarkable protein architecture has evolved to stabilize the human erythrocyte membrane and thereby enable mature cells to repeatedly withstand the shear stresses and deformations that are required as they traverse the capillaries in the peripheral circulation. Though the identity of the major proteins that comprise this architecture are known, atomic resolution structures of some isolated domains have been determined, and sparse data exist that suggest which amino acid residues are involved in the interactions, very little is currently known about the global structure of this complex of proteins. It is known that the cytoplasmic domain of the anion exchange protein (AE1; also known as band 3) serves as a critical organizing center for assembly of the major complex of proteins that stabilizes the membrane. It is also known that interactions between the cytoplasmic domain of band 3 (cdb), the membrane adaptor protein ankyrinR, and protein 4.2 are required for normal erythrocyte shape and membrane stability. Disruptions in this assembly lead to abnormally shaped erythroctyes and a condition known clinically as hemolytic anemia. Two problems that we have encountered at Vanderbilt is that DEER is not reliable for measurement of distances shorter than 20 ¿ and cw EPR is minimally sensitive to distances in the 18-22 ¿ range. Double Quantum Coherence (DQC), an alternative pulsed dipolar spectroscopy technique pioneered by the Freed group at ACERT fills in this gap very nicely and provides sensitivity to inter-probe distances to even shorter distances down to the 12 ¿ range. We will collaborate with the Center to make inter-probe distance measurements in the 12 to 20 ¿ range by DQC.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 一种非凡的蛋白质结构已经进化到稳定人类红细胞膜，从而使成熟细胞能够反复承受在外周循环中穿过毛细血管时所需的剪应力和变形。尽管组成这种结构的主要蛋白质的身份是已知的，一些孤立结构域的原子分辨结构已经确定，并且存在稀少的数据表明哪些氨基酸残基参与了相互作用，但目前对这种蛋白质复合体的全球结构知之甚少。众所周知，阴离子交换蛋白(AE1；也称为带3)的细胞质结构域是稳定膜的主要蛋白质复合体组装的关键组织中心。我们还知道，带3的细胞质结构域(CDB)、膜接头蛋白ankyrinR和蛋白4.2之间的相互作用是正常的红细胞形状和膜稳定性所必需的。这种组装的破坏会导致红血球畸形和一种临床上称为溶血性贫血的情况。我们在Vanderbilt遇到的两个问题是，鹿在测量短于20？的距离时不可靠，而CW EPR对18-22？范围内的距离最不敏感。双量子相干(DQC)是由Acert的FREED小组开创的一种替代脉冲偶极光谱技术，它很好地填补了这一空白，并提供了对探头间距离的敏感性，甚至可以精确到12？范围内的更短距离。我们将与该中心合作，由DQC进行12到20？范围内的探头间距离测量。