Novel Locations; Familiar Functions: Obscurin at the Cardiac Intercalated Disc

新地点；

基本信息

批准号：
8425488
负责人：
Maegen A. Borzok
金额：
$ 12.83万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-05 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8425488
关键词：
Acute Address Affinity Animals Ankyrins Architecture Binding Binding Sites Biochemical Biological Assay Biology Cardiac Cardiac Myocytes Cells Cellular Membrane Collaborations Complex Coupling Custom Cytoskeletal Proteins Cytoskeleton Data Development Development Plans Disease Disease model Dot Immunoblotting Echocardiography Elements Ensure Equipment Event Family Figs - dietary Fluorescence Microscopy Functional disorder Gene Delivery Genes Goals Grant Growth Health Heart Heart Diseases Heart failure Immunologic Techniques In Vitro Individual Injection of therapeutic agent Intercalated disc Intercellular Junctions Laboratories Lead Length Ligands Lipids Location Maintenance Mass Spectrum Analysis Mechanics Mediating Membrane Membrane Microdomains Mentors Methodology Molecular Molecular Biology Techniques Mus Muscle Muscle Cells Mutation Myocardial Infarction Normalcy PH Domain Pathology Performance Pericardial body location Phase Phosphorylation Phosphotransferases Physiological Physiology Play Positioning Attribute Protein Family Protein Isoforms Proteins Proteome Proteomics RNA Splicing Reagent Regulatory Pathway Relative (related person)Research Research Personnel Resources Role Sarcolemma Sarcomeres Sarcoplasmic Reticulum Signal Transduction Striated Muscles Subcellular structure Surface Plasmon Resonance Techniques Testing Time Tissues Training Training Support Translations United States National Institutes of Health Viral Work base career development cell motility design and construction experience genetic manipulation heart cell heart function in vivo innovation member mouse model mutant novel obscurin overexpression postnatal pressure research and development research study rho guanine nucleotide exchange factor p115 translational approach transmission process two-dimensional

项目摘要

Obscurins, cytoskeletal proteins discovered about a decade ago, are encoded by the single OBSCN gene and have been primarily studied in vertebrate striated muscles. The giant (~720-900 kDa) obscurin isoforms intimately surround sarcomeres at M-bands and Z-disks, where obscurins participate in their assembly, stability, and integration with other sarcomeric elements. The PI and her primary mentor (Dr. Kontrogianni- Konstantopoulos) are at the forefront of characterizing this novel family of proteins. Recently, the PI has identified a novel obscurin isoform (obscD), resulting from complex differential splicing at the 3' end of the gene that preferentially concentrates at the intercalated disc (ID) of cardiomyocytes. The ID is a unique membrane microdomain that mediates mechanical and electrical coupling between neighboring cells, allowing the synchronous beating of the heart. Importantly, mutations and disruptions of ID proteins interrupt regular heart function, consequently leading to heart disease. Consistent with its ID localization,biochemical assays indicate that obscD exists in a complex with other proteins of the ID proteome. Furthermore, preliminary proteomic analysis has indicated that obscD is regulated via phosphorylation, and its phosphorylation state is altered in the dystrophic disease model. This project aims to elucidate the function that obscD plays at the ID and determine the role of obscD's phosphorylation in physiology and pathophysiology. The PI has assembled an accomplished individualized mentoring committee who will provide the necessary training and support to accomplish the proposed research, as well as facilitate the growth of the PI. The PI will remain at UM SOM, working under the guidance of Drs. Kontrogianni-Konstantopoulos, Rogers, and Stanley. Additionally, the PI will also benefit from the enrichment of Dr. Sellers, at nearby NIH. Through this mentoring team and the collaborations set up by the PI, she will have access to both routine and state-of-the-art equipment and reagents necessary to successfully and efficiently complete the proposed research. In addition, the PI will have excellent resources for career development at the UM SOM and NIH campuses. The proposed research and career development plan will enable the PI to investigate the phosphorylation of obscD and elucidate its in vivo physiological and pathophysiological function at the ID. Furthermore, they will facilitate her development as a diversified independent muscle biology investigator who integrates traditional biochemical and molecular biology techniques with whole tissue translational approaches.

Obscurins 是大约十年前发现的细胞骨架蛋白，由单个 OBSCN 基因编码，主要在脊椎动物横纹肌中进行研究。巨大的（~720-900 kDa）暗蛋白亚型紧密围绕 M 带和 Z 盘的肌小节，暗蛋白在其中参与其组装、稳定性以及与其他肌节元件的整合。 PI 和她的主要导师（Kontrogianni-Konstantopoulos 博士）处于表征这一新型蛋白质家族的最前沿。最近，PI 发现了一种新的 obscurin 异构体 (obscD)，该异构体是由该基因 3' 端的复杂差异剪接产生的，该基因优先集中在心肌细胞的闰盘 (ID) 处。 ID 是一种独特的膜微域，可介导相邻细胞之间的机械和电耦合，从而实现心脏的同步跳动。重要的是，ID 蛋白的突变和破坏会扰乱正常的心脏功能，从而导致心脏病。与 ID 定位一致，生化分析表明 obscD 与 ID 蛋白质组的其他蛋白质存在于复合物中。此外，初步蛋白质组学分析表明obscD通过磷酸化进行调节，并且其磷酸化状态在营养不良性疾病模型中发生改变。该项目旨在阐明obscD在ID中发挥的功能，并确定obscD磷酸化在生理学和病理生理学中的作用。 PI 组建了一个完善的个性化指导委员会，该委员会将提供必要的培训和支持以完成拟议的研究，并促进 PI 的成长。 PI 将继续留在 UM SOM，在 Drs. 的指导下工作。康特罗贾尼-康斯坦托普洛斯、罗杰斯和斯坦利。此外，PI 还将受益于附近 NIH 的 Sellers 博士的丰富经验。通过这个指导团队以及 PI 建立的合作，她将能够获得成功高效完成拟议研究所需的常规和最先进的设备和试剂。此外，PI 将在 UM SOM 和 NIH 校区拥有出色的职业发展资源。拟议的研究和职业发展计划将使 PI 能够研究 obscD 的磷酸化并阐明其在 ID 的体内生理和病理生理功能。此外，它们将促进她发展成为一名多元化的独立肌肉生物学研究者，将传统的生化和分子生物学技术与全组织转化方法相结合。