Assembly of Spectrin Isoforms

血影蛋白亚型的组装

• 批准号: 7350729
• 负责人: LESLIE W. FUNG
• 金额: $ 2.9万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7350729
• 关键词: Affect; Affinity; Alzheimer' s Disease; Anemia, Hemolytic, Congenital; Antibodies; Biological Models; Brain; Cell physiology; Class; Complex; Diagnosis; Dimerization; Disease; Disease Marker; Drug Delivery Systems; Ensure; Erythrocytes; Erythroid; Exhibits; Hematological Disease; Individual; Information Systems; Membrane; Methods; Modeling; Molecular; Molecular Conformation; Numbers; Patients; Peptides; Physiology; Play; Property; Protein Isoforms; Proteins; Proteomics; Range; Role; Saccharomyces; Screening procedure; Site; Skeletal system; Spectrin; Spin Labels; Structure; System; Testing; base; brain cell; cell type; design; insight; interest; member; nervous system disorder; prevent; protein protein interaction; recombinant peptide

Our broad, long-term objectives are to understand conformations of critical regions in various spectrin isoforms and to correlate specific structural features in these isoforms with functions unique to individual isoforms. We begin by studying the association affinities and local structures at the tetramerization site of two different spectrins, using recombinant peptide model systems. The four model peptides to be used are Spod-1-368 (erythrocyte), Spodl-1-359 (brain), Spl31-1689-2083 (erythrocyte) and Spl311-1697-2091 (brain). We have already determined the junction region conformation of Spod-1-368. The junction region conformations of the other three peptides will be determined in this project by site-directed spin labeling EPR methods. Other biophysical methods will be used to ensure the integrity of the peptides. The association affinity will also be determined to correlate with the junction structural properties in these peptides. Alternate studies are proposed if the first part of this project shows that the junction regions play no role in regulating association affinities. The specific aims are (1) To characterize the junction region conformation for Spodl-1-359, (2) To determine association affinities of Spod-1-368 and Spod11-359 with SPl31-1689-2083 and with Spl311-1697-2091, (3) To characterize the junction region conformations in Sp131-1689-2083 and in Spl311-1697-2091, (4) To characterize the or- and D-junction region conformations and interactions in the _13complex for both _1131and _11131s1ystems, and (5) To identify key residues in Sp_11-1-359 that affect its association affinity with Spl31-1689-2083 and with Spl311-1697-2091. Findings from these studies will provide insight toward developing molecular understanding of normal physiology involving spectrin molecules in erythrocytes, brain cells and other types of cells, and of diseases related to spectrin tetramers, such as hereditary hemolytic anemia, and neurological disorders. We believe that new disease markers and drug targets can be identified to help design products to prevent, diagnose and treat diseases through functional proteomic studies of spectrin isoforms and their protein-protein interactions. In addition to the spectrin systems, the information obtained from this study will also provide insight toward a better understanding of naturally occurring coiled-coil subunit-subunit associations.

我们广泛的长期目标是了解各种关键区域的构象 光谱同工型，并将这些同工型中的特定结构特征与独特的功能相关联 单个同工型。我们首先研究关联的亲和力和本地结构 使用重组肽模型系统的两个不同光谱的四聚体位点。四个模型 要使用的肽是Spod-1-368（红细胞），Spodl-1-359（Brain），SPL31-1689-2083（Erythrocyte）和 SPL311-1697-2091（大脑）。我们已经确定了Spod-1-368的连接区构象。 其他三个肽的交界区域构象将在该项目中确定 站点定向的自旋标记EPR方法。其他生物物理方法将用于确保 肽。联想亲和力还将确定与结构相关 这些肽中的性质。如果该项目的第一部分表明，则提出了替代研究 交界区域在规范关联亲和力中没有作用。具体目的是（1）表征 Spodl-1-359，（2）的连接区域构象，以确定Spod-1-368和 SPOD11-359，具有SPL31-1689-2083和SPL311-1697-2091，（3），以表征交界区域 SP131-1689-2083和SPL311-1697-2091中的构象，（4）表征OR-和D-JENCTION区域 _1131和_11131S1ystems和（5）识别密钥 SP_11-1-359中的残留物影响其与SPL31-1689-2083和SPL311-1697-2091的关联亲和力。 这些研究的发现将为建立对正常的分子理解提供洞察力 涉及红细胞，脑细胞和其他类型细胞中的光谱分子的生理学以及疾病 与光谱四聚体有关，例如遗传性溶血性贫血和神经系统疾病。我们相信 可以确定新的疾病标志物和药物靶标，以帮助设计产品以防止，诊断 并通过光谱同工型的功能蛋白质组学研究及其蛋白质蛋白质治疗疾病 互动。除了光谱系统外，从这项研究中获得的信息还将提供 洞悉对自然发生的盘绕蛋白亚基 - 亚基的关联的洞察力。