Structural studies of a spectrin/ankyrin complex

血影蛋白/锚蛋白复合物的结构研究

• 批准号: 8197746
• 负责人: Alfonso Mondragon
• 金额: $ 27.8万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8197746
• 关键词: Adaptor Signaling Protein; Affinity; Amino Acids; Anions; Ankyrin Repeat; Ankyrins; Architecture; Arrhythmia; Binding; Binding Sites; Biochemical; Biological; Blood capillaries; C-terminal; Calorimetry; Cardiac; Cardiac Death; Cell membrane; Cell physiology; Cellular Structures; Clinical Data; Complex; Cytoskeleton; Death Domain; Defect; Disease; Elements; Erythrocytes; Erythroid; Goals; Hematological Disease; Hereditary Elliptocytosis; Hereditary Spherocytosis; Human; In Vitro; Individual; Infection; Knowledge; Long QT Syndrome; Malaria; Maps; Measures; Membrane; Methods; Molecular; Morphology; Mutation; Normal Cell; Personal Satisfaction; Process; Proteins; Resolution; Roentgen Rays; Role; Shapes; Spectrin; Structure; Surface Plasmon Resonance; Techniques; Testing; Work; X-Ray Crystallography; analytical ultracentrifugation; base; capillary; complex R; human disease; in vivo; insight; interest; link protein; mutant; protein complex; public health relevance; research study; three dimensional structure

DESCRIPTION (provided by applicant): In erythrocytes, the reversible deformability needed to accomplish passage through capillaries is largely controlled by a vast network of proteins attached to the membrane. Spectrin is a key element of this network. The attachment between the membrane and the spectrin cytoskeleton is accomplished through an adaptor protein, ankyrin, which bridges the interaction between 2-spectrin and the membrane bound anion- exchange transporter band 3. The ankyrin-spectrin interaction has important implications for normal cell function and for some diseases. Hereditary spherocytosis and hereditary elliptocytosis are human blood disorders where mutations have been mapped to spectrin and ankyrin. Ankyrin defects are also responsible for the cardiac death syndrome, long-QT cardiac arrhythmia. In addition, ankyrin, and in particular its spectrin binding site, has been involved in malaria red blood cell infection. This proposal is concerned with biochemical and structural studies of human erythroid 2-spectrin and ankyrin with the goal of providing an atomic understanding of this complex. In the past few years we have made substantial progress towards this goal, including solving the atomic structure of the individual binding domains of 2-spectrin and ankyrin R, and the complex formed by these two domains. These structures are starting to provide important structural information on these two critical molecules and allowing us to relate the structure to the wealth of existing functional, biochemical, and clinical data. For the next project period we propose to continue and expand our studies of ankyrin and spectrin. The specific aims for this proposal are: 1) to continue our work on the three dimensional structure of a complex of the spectrin binding domain of ankyrin and the ankyrin binding domain of spectrin and to study the role of different amino acids and different regions in the spectrin/ankyrin recognition process and, 2) to expand our biophysical and structural studies to include other regions of spectrin and ankyrin that have not been characterized structurally. The work is based on a combination of molecular biological, biophysical, and biochemical methods to produce and characterize the molecules that we require for our work, X-ray crystallography to solve their atomic structures, and cellular studies to correlate our in vitro findings to observations in erythrocytes. The knowledge of the structure of the complex formed by spectrin and ankyrin promises to provide important and relevant information on this critical protein complex involved in a central cellular process and with important implications to human well-being. PUBLIC HEALTH RELEVANCE: We propose to study and characterize the structure of human 2I-spectrin and ankyrin R, whose role is to attach the red blood cell membrane to the spectrin cytoskeleton. Defects in these proteins are linked to two blood disorders, hereditary spherocytosis and hereditary elliptocytosis. In addition, ankyrin defects are involved in long-QT cardiac arrhythmia and other cardiac problems.

描述（由申请人提供）：在红细胞中，完成通过毛细血管所需的可逆变形性在很大程度上由附在膜上的大量蛋白质网络控制。 Spectrin是该网络的关键要素。膜与光谱细胞骨架之间的附着是通过衔接蛋白Ankyrin完成的，arnkyrin桥接了2谱蛋白与膜结合的阴离子交换转运蛋白带3之间的相互作用。Ankyrin-spectrin spectrin相互作用对正常细胞功能具有重要意义，对正常细胞功能具有重要意义。遗传性球细胞增多症和遗传性椭圆细胞增多症是人类血液疾病，其中突变已被映射到谱蛋白和脚踝蛋白。 Ankyrin缺陷还导致心脏死亡综合征长QT心律不齐。另外，氨基林，尤其是其光谱结合位点，还参与了疟疾红细胞感染。 该建议涉及人类红细胞2-谱蛋白和Ankyrin的生化和结构研究，目的是提供对这一复合物的原子学理解。在过去的几年中，我们取得了重大进展，包括解决2-谱蛋白和Ankyrin R的单个结合域的原子结构，以及由这两个域形成的复合物。这些结构开始提供有关这两个关键分子的重要结构信息，并允许我们将结构与现有功能，生化和临床数据的财富联系起来。在下一个项目期间，我们建议继续并扩大对弯蛋白和光谱蛋白的研究。该提案的具体目的是：1）继续我们在安氨基蛋白的谱蛋白结合结构域的三维结构上继续我们的工作结构上。这项工作基于分子生物学，生物物理和生化方法的结合，以产生和表征我们工作所需的分子，X射线晶体学以求解其原子结构以及将我们的体外发现与红细胞观测结果相关的细胞研究。 Spectrin和Ankyrin形成的复合物结构的知识有望提供有关此关键蛋白质复合物与中央细胞过程有关的重要和相关信息，并且对人类的幸福感具有重要意义。 公共卫生相关性：我们建议研究和表征人2-specrin和Ankyrin R的结构，它们的作用是将红细胞膜连接到谱蛋白细胞骨架上。这些蛋白质中的缺陷与两种血液疾病，遗传性球细胞增多症和遗传椭圆细胞增多症有关。此外，雄性心律失常和其他心脏问题涉及脚踝缺陷。