PROTEOMIC ANALYSES OF PERLECAN MRNA-ASSOCIATED PROTEIN COMPLEXES

PERLECAN mRNA 相关蛋白复合物的蛋白质组学分析

• 批准号: 8365859
• 负责人: CARLOS Benjamin HIRSCHBERG
• 金额: $ 1.28万
• 依托单位: UNIVERSITY OF WASHINGTON
• 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 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365859
• 关键词: Affinity Chromatography; Alternative Splicing; Animals; Arthritis; Atherosclerosis; Basement membrane; Biology; Development; Diabetes Mellitus; Foundations; Funding; Fungal Genome; Grant; Heparan Sulfate Proteoglycan; Link; Longitudinal Studies; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; National Center for Research Resources; Pathologic Processes; Principal Investigator; Proteins; Proteomics; RNA Splicing; Regulation; Research; Research Infrastructure; Resources; Source; United States National Institutes of Health; cost; insight; mRNA Precursor; perlecan; protein complex; research study

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. Perlecan is a major component of heparan sulfate proteoglycans in basement membrane, which is widely expressed in animal bodies. Its expression is dynamically regulated during development and its aberrant expression is linked to many pathological processes, such as diabetes, atherosclerosis, arthritis and cancer. However, it has been poorly understood how the expression of perlecan is properly regulated. Here we show that by affinity purification we were able to obtain two protein complexes that are specifically associated with perlecan mRNA, likely involved in its pre-mRNA splicing. We will examine the purified protein complexes by mass spectrometry and analyze the proteins for their functional significances in both constitutive and alternative splicing. The proposed experiments will provide mechanistic insights into the regulation of constitutive and/or alternative splicing of perlecan pre-mRNA, which will establish a foundation for long-term studies to elucidate the mechanism of alternative splicing in regulating the expression of perlecan.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 Perlecan是地下膜中硫酸乙酰肝素蛋白聚糖的主要组成部分，该膜在动物体中广泛表达。它的表达在发育过程中受动态调节，其异常表达与许多病理过程有关，例如糖尿病，动脉粥样硬化，关节炎和癌症。但是，众所周知，perlecan的表达是如何适当调节的。在这里，我们表明，通过亲和力纯化，我们能够获得两个与perlecan mRNA特别相关的蛋白质复合物，这些蛋白可能与其MRNA剪接有关。我们将通过质谱检查纯化的蛋白质复合物，并分析蛋白质在本构和替代剪接中的功能意义。提出的实验将为perlecan前MRNA的构成和/或替代剪接的调节提供机械见解，该剪接将建立长期研究的基础，以阐明在调节perlecan表达时替代剪接机制。