首页> 外文学位 >Identification of novel protein-protein interactions and functional analysis in the mouse photoreceptor of the hypothetical protein FLJ33282-Als2cr4 and the small Rho GTPase, Rnd2.
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Identification of novel protein-protein interactions and functional analysis in the mouse photoreceptor of the hypothetical protein FLJ33282-Als2cr4 and the small Rho GTPase, Rnd2.

机译:假设蛋白FLJ33282-Als2cr4和小的Rho GTPase Rnd2的小鼠光感受器中新型蛋白-蛋白相互作用的鉴定和功能分析。

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摘要

The main focus of this research project was to decipher the role of the hypothetical protein FLJ33282-Als2cr4 (amyotrophic lateral sclerosis 2 [juvenile] chromosome region, candidate 4; NM_001037812) in the mouse retina. We characterized its molecular structure, cellular interacting partners and potential biochemical functions. Previous in situ hybridization and gene expression profiles showed mRNAs encoding Als2cr4 are abundant in the eye, hippocampus, cerebellum, and olfactory bulb from embryonic day 15 and throughout adulthood in mouse.;Antibodies were developed from predicted antigenic epitopes of Als2cr4, characterized, and used to examine protein expression and morphological localization in retinas from light-adapted and dark-adapted mice with immunohistochemistry, immunoblot analysis and immuno-electron microscopy, followed with immunoprecipitation studies to identify potential functional interacting proteins by mass spectroscopy analysis.;My doctoral work demonstrates that Als2cr4 is a 45 kDa insoluble protein, which is highly enriched in mouse retina and localizes to photoreceptor outer segments, ciliary complex, and outer plexiform layer. The immuno-electron microscopy for Als2cr4 verifies its expression pattern in the discs of photoreceptor outer segments. Furthermore, immunoprecipitation analysis followed by mass spectroscopy identified eight novel interacting partners.;Our data provides evidence that the novel protein Als2cr4 has a predicted tetraspanin-like membrane structure, is localized to photoreceptors and to the post-synaptic outer plexiform layer, and interacts with cytoskeletal proteins. We predict that Als2cr4 is involved in membrane transport between the photoreceptor inner segment and outer segment and is a key component in maintaining outer segment structural integrity.
机译:该研究项目的主要重点是破译假设蛋白FLJ33282-Als2cr4(肌萎缩性侧索硬化2 [未成年]染色体区域,候选4; NM_001037812)的作用。我们表征了它的分子结构,细胞相互作用的伙伴和潜在的生化功能。先前的原位杂交和基因表达谱显示,从胚胎第15天开始到成年小鼠的整个眼睛,海马,小脑和嗅球中都编码Als2cr4的mRNA丰富;从预测的Als2cr4抗原表位开发了抗体,对其进行了表征和使用用免疫组织化学,免疫印迹分析和免疫电子显微镜检查光适应和黑暗适应小鼠视网膜中蛋白质的表达和形态学定位,然后进行免疫沉淀研究,通过质谱分析鉴定潜在的功能相互作用蛋白。 Als2cr4是一种45 kDa的不溶蛋白,在小鼠视网膜中高度富集,并位于感光器外部节段,睫状复合体和外部丛状层。 Als2cr4的免疫电子显微镜检查可验证其在感光器外部节段盘中的表达模式。此外,免疫沉淀分析和质谱分析确定了八个新的相互作用伙伴。我们的数据提供了证据,表明新蛋白Als2cr4具有预测的四跨膜蛋白样膜结构,位于光感受器和突触后外丛状层,并与细胞骨架蛋白。我们预测Als2cr4参与感光器内部部分和外部部分之间的膜运输,并且是维持外部部分结构完整性的关键组成部分。

著录项

  • 作者

    Zuniga, Freddi Isaac.;

  • 作者单位

    University of Southern California.;

  • 授予单位 University of Southern California.;
  • 学科 Biology Molecular.;Biology Neurobiology.;Chemistry Biochemistry.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 147 p.
  • 总页数 147
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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