Assembly of Spectrin Isoforms

血影蛋白亚型的组装

• 批准号: 7928385
• 负责人: LESLIE W. FUNG
• 金额: $ 16.18万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7928385
• 关键词: Affect; Affinity; Alzheimer' s Disease; Anemia, Hemolytic, Congenital; Antibodies; Biological Models; Brain; Cell physiology; Complex; Diagnosis; Dimerization; Disease; Disease Marker; Drug Delivery Systems; Ensure; Erythrocytes; Erythroid; Exhibits; Hematological Disease; Individual; Information Systems; Membrane; Methods; Modeling; Molecular; Molecular Conformation; Patients; Peptides; Physiology; Play; Property; Protein Isoforms; Proteins; Proteomics; Role; Saccharomyces; Screening procedure; Site; Spectrin; Spin Labels; Structure; System; Testing; base; brain cell; cell type; design; insight; interest; member; nervous system disorder; prevent; protein protein interaction; recombinant peptide; skeletal

DESCRIPTION (provided by applicant): 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 Spalpha-1-368 (erythrocyte), SpalphaII-1-359 (brain), SpbetaI-1689-2083 (erythrocyte) and SpbetaII-1697-2091 (brain). We have already determined the junction region conformation of SpalphaI-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 Spalphall-1-359, (2) To determine association affinities of Spalpha-1-368 and SpalphaII-1-359 with SpbetaI-1689-2083 and with SpbetaII-1697-2091, (3) To characterize the junction region conformations in SpbetaI-1689-2083 and in SpbetaII-1697-2091, (4) To characterize the alpha- and beta-junction region conformations and interactions in the alphabeta complex for both alphaIbetaI and alphaIIbetaII systems, and (5) To identify key residues in SpalphaII-1-359 that affect its association affinity with SpbetaI-1689-2083 and with SpbetaII-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.

描述（由申请人提供）：我们广泛的长期目标是了解各种光谱同工型中关键区域的构象，并将这些同工型中的特定结构特征与单个同工型独有的功能相关联。我们首先使用重组肽模型系统研究两个不同光谱的四聚体位点的关联亲和力和局部结构。要使用的四种模型肽是spalpha-1-368（erythrocyte），spalphaii-1-359（脑），SPBETAI-1689-2083（erythrocyte）和SPBETAIII-1697-2091（Brain）。我们已经确定了Spalphai-1-368的连接区域构象。其他三个肽的连接区域构象将通过位置定向的自旋标记EPR方法确定。其他生物物理方法将用于确保肽的完整性。还将确定关联亲和力与这些肽中的连接结构特性相关。如果该项目的第一部分表明，交界处地区在调节关联亲和力中没有作用，则提出了替代研究。 （1）的特定目的是表征spalphall-1-359，（2）的连接区域构型，以确定spalpha-1-368和spalphaii-1-359与SPBETAI-1689-2083的关联相关性，以及SPBETAII-1697-2091，（3）SPBEAI 33 3和SPBIAIN 30 3和3号纽约。 SPBETAII-1697-2091，（4）为Alphaibetai和Alphaiibetaii Systems的Alphabeta复合体中的α-和β结区构象和相互作用，以及（5），以识别Spalphaii-1-359中与Spalphaii-1-359的关键残基，这些残基影响其与Spbbetai-16-16-16-1683的关联， SPBETAII-1697-2091。 这些研究的发现将为对涉及红细胞，脑细胞和其他类型细胞的光谱分子的正常生理学的分子理解提供洞察力，以及与谱蛋白四聚体有关的疾病，例如遗传性造血性疾病贫血和神经学障碍。我们认为，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，诊断和治疗疾病，通过谱蛋白同工型的功能性蛋白质组学研究及其蛋白质蛋白质相互作用。除Spectrin系统外，从这项研究获得的信息还将提供洞察力，以更好地理解天然发生的盘绕圈亚基 - 亚基相关性。

项目成果

Nanomole-scale protein solid-state NMR by breaking intrinsic 1HT1 boundaries.

• DOI: 10.1038/nmeth.1300
• 发表时间: 2009-03
• 期刊: Nature methods
• 影响因子: 48
• 作者: Wickramasinghe NP;Parthasarathy S;Jones CR;Bhardwaj C;Long F;Kotecha M;Mehboob S;Fung LW;Past J;Samoson A;Ishii Y
• 通讯作者: Ishii Y

Association studies of erythroid alpha-spectrin at the tetramerization site.

红系α-血影蛋白四聚位点的关联研究。

• DOI: 10.1111/j.1365-2141.2009.07876.x
• 发表时间: 2009
• 期刊: British journal of haematology
• 影响因子: 6.5
• 作者: Lam,VinhQ;Antoniou,Chloe;Rolius,Ramunas;Fung,LeslieW-M
• 通讯作者: Fung,LeslieW-M

Inhibition of calpain but not caspase activity by spectrin fragments.

血影蛋白片段抑制钙蛋白酶但不抑制半胱天冬酶活性。

• DOI: 10.2478/s11658-010-0015-3
• 发表时间: 2010
• 期刊: Cellular & molecular biology letters
• 影响因子: 8.3
• 作者: Rolius,Ramunas;Antoniou,Chloe;Nazarova,LidiaA;Kim,StephenH;Cobb,Garrett;Gala,Pooja;Rajaram,Priyanka;Li,Qufei;Fung,LeslieW-M
• 通讯作者: Fung,LeslieW-M

Brain proteins interacting with the tetramerization region of non-erythroid alpha spectrin.

• DOI: 10.2478/s11658-007-0028-8
• 发表时间: 2007
• 期刊: Cellular & molecular biology letters
• 影响因子: 8.3
• 作者: Oh Y;Fung LW
• 通讯作者: Fung LW

Yeast two-hybrid and itc studies of alpha and beta spectrin interaction at the tetramerization site.

酵母双杂交和 itc 研究四聚位点 α 和 β 血影蛋白相互作用。

• DOI: 10.2478/s11658-011-0017-9
• 发表时间: 2011
• 期刊: Cellular & molecular biology letters
• 影响因子: 8.3
• 作者: Sevinc,Akin;Witek,MartaA;Fung,LeslieW-M
• 通讯作者: Fung,LeslieW-M