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

新地点；

• 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
• 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

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.

大约十年前发现的近距离细胞骨架蛋白是由单个淫秽基因编码的，并且主要在脊椎动物横纹肌肉中进行了研究。巨型（〜720-900 kDa）遮盖的同工型紧密围绕M频段和Z-Disks的肉瘤，在该肉瘤中，遮盖蛋白参与了它们的组装，稳定性，并与其他肉瘤元素集成。 PI和她的主要导师（Kontrogianni-Konstantopoulos博士）正处于描述这种新型蛋白质家族的最前沿。 最近，PI鉴定出一种新型的模糊同工型（BIBSD），这是由于基因3'末端的复杂差剪接而产生的，该剪接优先集中在心肌细胞的插入式椎间盘（ID）上。 ID是一个独特的膜微区域，可介导相邻细胞之间的机械和电耦合，从而使心脏同步跳动。重要的是，ID蛋白的突变和破坏中断常规心脏功能，因此导致心脏病。与其ID定位一致，生化测定法表明，与ID蛋白质组的其他蛋白质的复合物存在于复合物中。此外，初步的蛋白质组学分析表明，淫秽通过磷酸化调节，其磷酸化状态在营养不良疾病模型中改变了。该项目旨在阐明淫秽对ID的功能，并确定淫秽磷酸化在生理和病理生理学中的作用。 PI已组建了一个成熟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的增长。在Drs的指导下，PI将留在UM SOM。 Kontrogianni-Konstantopoulos，Rogers和Stanley。此外，PI还将受益于附近NIH的Sellers博士的丰富。通过这个指导团队以及PI建立的合作，她将可以使用常规和最先进的设备以及成功，有效地完成拟议的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校园内拥有出色的资源。拟议的研究和职业发展计划将使PI能够研究obsd的磷酸化，并在ID处阐明其体内生理和病理生理功能。此外，他们将促进她作为一名多元化的独立肌肉生物学研究者的发展，他们将传统的生化和分子生物学技术与整个组织转化方法相结合。