COBRE: OUHSC: PROTEOMICS/BIOINFORMATICS CORE

COBRE：OUHSC：蛋白质组学/生物信息学核心

• 批准号: 7959973
• 负责人: HIROYUKI MATSUMOTO
• 金额: $ 6.36万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959973
• 关键词: 4 hydroxynonenal; Actins; Affinity; Analytical Biochemistry; Area; Binding; Biochemical; Bioinformatics; Caveolae; Centers of Research Excellence; Complex; Computer Retrieval of Information on Scientific Projects Database; Databases; Detergents; Development; Diabetic Retinopathy; Digestion; Funding; Gel; Goals; Grant; Image; Immunoblot Analysis; Institution; Insulin Receptor; Journals; Knockout Mice; LIF gene; Light; Mass Spectrum Analysis; Membrane; Mentors; Modification; Mus; Oklahoma; Oxidative Stress; Oxygen; PTPN1 gene; Paper; Pathologic Processes; Photoreceptors; Pigment Epithelium; Protein Tyrosine Phosphatase; Proteins; Proteome; Proteomics; Publications; Receptor Signaling; Research; Research Personnel; Resistance; Resources; Retina; Retinal; Retinal Cone; Retinal Degeneration; Retinal Diseases; Retinal Photoreceptors; Retinol dehydrogenase; Rod Outer Segments; Role; Source; Stress; Structure of retinal pigment epithelium; System; Transgenic Mice; Trypsin; Two-Dimensional Gel Electrophoresis; Tyrosine; United States National Institutes of Health; Vision research; Work; adduct; cyclic-nucleotide gated ion channels; human RIPK1 protein; leukemia inhibitory factor; mouse model; neurochemistry; novel; oxidation; photoreceptor degeneration

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 1) The John D. Ash (ECI) Project: The primary project was to characterize the proteome of mouse retinas in development and in transgenic mice expressing leukemia inhibitory factor. A secondary project was to characterize the retina proteome of the retina in a mouse model of oxygen induced retinopathy. A plan to use this facility to identify proteins modified by oxidation including 4-HNE, and nitro-tyrosine is due next year. The goal of the study is to determine whether LIF protects photoreceptors by reducing global or specific protein oxidation or promoting protection despite protein oxidation. 2) The Raju V. Rajala (ECI) Project: This project aims to investigate proteins which interact with the retinal insulin receptor. As a result of our effort we have successfully identified actin as one of the retinal insulin receptor interacting proteins. This finding opens a new area of research to study the actin-insulin receptor interactions in diseased states such as diabetic retinopathy and stress-induced retinal degeneration. Another project within this context is to identify novel proteins which bind to p85 subunit of PI3K. We also aim to identify substrates to PTP1B protein tyrosine phosphatase. The results will significantly contribute to the understanding of the retinal insulin receptor signaling. 3) The Xi-Qing Ding (PJI) Project: Photoreceptor cyclic nucleotide-gated (CNG) channel complex assembly and interaction with adjacent proteins are crucial for the channel function. We plan to identify the modulatory molecules of cone CNG channel using a proteomics approach. Affinity purified channel complexes are analyzed by the proteomics/analytical biochemical approaches. This includes one- and two- dimensional gel electrophoresis, in-gel trypsin digestion, mass spectrometry and MS/MS analysis, and database searching. 4) The Michael H. Elliott (PJI) Project: We have made moderate/heavy use of the Proteomics/Analytical Biochemistry Module in the past funding period, which will continue next year. We have applied a mass spectrometric analysis to examine the protein composition of detergent-resistant membranes from photoreceptor rod outer segments using the Proteomics Module and this work resulted in a 2008 publication in the Journal of Neurochemistry. In addition, the Li-Cor Odyssey IR Imaging system was essential for quantitative immunoblot analysis in this paper and in another study. In the upcoming grant period, we plan to use the Proteomics/Analytical Biochemistry Module to identify protein targets that are present in caveolae isolated from retinal pigment epithelium. The results will significantly contribute to a better understanding of the role of pigment epithelium for keeping the functional integrity of retinal photoreceptor cells. 5) Anne Kasus-Jacobi (PJI) Project: We plan to use a proteomic approach 1) to Identify oxidative modification of the retinol dehydrogenase RDH12 in mouse retina, during light-induced oxidative stress, and 2) to identify the proteins forming Michael's adducts with 4-Hydroxynonenal in wild-type and Rdh12 knockout mouse retinas, during light-induced oxidative stress. The results to be obtained from these specific aims will reveal the pathological process during the light-induced oxidative stress, which will shed light to the understanding of photoreceptor degeneration.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 1)约翰D.灰（ECI）项目： 主要项目是表征发育中的小鼠视网膜和表达白血病抑制因子的转基因小鼠的蛋白质组。第二个项目是表征氧诱导视网膜病变小鼠模型中视网膜的视网膜蛋白质组。 一个计划，使用这个设施，以确定蛋白质修饰的氧化，包括4-HNE，和硝基酪氨酸是由于明年。该研究的目的是确定LIF是否通过减少整体或特定蛋白质氧化或促进蛋白质氧化的保护来保护光感受器。 2)Raju V. Rajala（ECI）项目： 该项目旨在研究与视网膜胰岛素受体相互作用的蛋白质。由于我们的努力，我们已经成功地确定肌动蛋白作为视网膜胰岛素受体相互作用的蛋白质之一。这一发现开辟了一个新的研究领域，研究肌动蛋白-胰岛素受体在疾病状态下的相互作用，如糖尿病视网膜病变和应激诱导的视网膜变性。 在此背景下的另一个项目是鉴定与PI 3 K的p85亚基结合的新蛋白。我们还旨在确定底物的PTP 1B蛋白酪氨酸磷酸酶。这一结果将有助于对视网膜胰岛素受体信号转导机制的理解。 3)Xi-Qing Ding（PJI）项目： 光感受器环核苷酸门控（CNG）通道复合物的组装和与相邻蛋白质的相互作用对通道功能至关重要。我们计划使用蛋白质组学方法鉴定锥CNG通道的调节分子。 通过蛋白质组学/分析生物化学方法分析亲和纯化的通道复合物。这包括一维和二维凝胶电泳、凝胶内胰蛋白酶消化、质谱和MS/MS分析以及数据库搜索。 4)迈克尔·H。Elliott（PJI）项目： 在过去的资助期内，我们适度/大量使用蛋白质组学/分析生物化学模块，明年将继续使用。我们已经应用了质谱分析，以检查使用蛋白质组学模块的感光杆外节的抗洗涤剂膜的蛋白质组成，这项工作导致在2008年发表在神经化学杂志。此外，Li-Cor Odyssey IR成像系统对于本文和另一项研究中的定量免疫印迹分析至关重要。在即将到来的资助期内，我们计划使用蛋白质组学/分析生物化学模块来鉴定从视网膜色素上皮分离的小窝中存在的蛋白质靶点。这一结果将有助于更好地理解色素上皮在维持视网膜感光细胞功能完整性中的作用。 5)Anne Kasus-Jacobi（PJI）项目： 我们计划使用蛋白质组学方法1）鉴定在光诱导的氧化应激期间小鼠视网膜中视黄醇脱氢酶RDH 12的氧化修饰，和2）鉴定在光诱导的氧化应激期间野生型和Rdh 12敲除小鼠视网膜中与4-羟基壬烯醛形成迈克尔加合物的蛋白质。这些研究结果将揭示光诱导的氧化应激过程中的病理过程，从而为理解光感受器变性提供线索。