DENNIS POST-DOC/TECHNICIAN SUPPORT

丹尼斯博士后/技术员支持

• 批准号: 8168294
• 负责人: Douglas Dennis
• 金额: $ 6.45万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168294
• 关键词: Atomic Force Microscopy; Biogenesis; Cell membrane; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Drug Delivery Systems; Electron Microscopy; Environment; Fluorescence; Funding; Genetic; Goals; Grant; Institution; Knowledge; Medical; Membrane; Modeling; Movement; Polymers; Postdoctoral Fellow; Process; Protein Binding; Proteins; Proteomics; Research; Research Personnel; Resources; Source; Structure; Subcellular Fractions; Thick; United States National Institutes of Health; Universities; Western Blotting; periplasm; programs

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. This proposal accomplishes the AREA program objectives of: 1) supporting meritorious research; 2) exposing undergraduates to research; and 3) strengthening the research environment in non-research intensive universities. The goal of this research is to elucidate the mechanism of polyhydroxyalkanoate inclusion biogenesis. Electron microscopy studies have been unable to resolve the structure of PHA inclusions and this has inhibited movement toward a cohesive model of inclusion biogenesis. Employing atomic force microscopy, we have determined that there are three layers of structure, an outer envelope that is the thickness of a membrane bilayer, a middle network layer, and an underlying crystalline lamellar layer. Genetic studies have indicated that the middle network is comprised at least partially of PhaP and that PhaP is likely to be translocated to the periplasm. Thus, it would appear that inclusion biogenesis may occur by movement of protein and/or proteins to the periplasm and budding through the cytoplasmic membrane into the cytoplasm, facilitating the acquisition of the cytoplasmic membrane as an envelope. The goal of this research is to prove or disprove this supposition. The specific aims of the research are: 1) definitively prove periplasmic localization of PhaP via fluorescence localization and Western blot analyses of subcellular fractions, 2) demonstrate that the inclusion envelope is derived from the cytoplasmic membrane by proteomic analysis, and 3) characterize proteins that bind transiently and permanently to PhaP in hopes of elucidating the mechanism of inclusion biogenesis. Ultimately, the goal of the research is to enlarge our knowledge of inclusion biogenesis to the point that this process can be controlled and utilized for medical applications. For instance, it could be envisioned that instead of polymer being inserted into the inclusion, bioactive compounds could be inserted, making the inclusion into a drug delivery vehicle.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该提案实现了以下方面的区域计划目标：1）支持有罪研究； 2）使大学生接受研究； 3）加强非研究密集大学的研究环境。这项研究的目的是 阐明多羟基烷烃包容生物发生的机制。电子显微镜研究无法解析PHA夹杂物的结构，这抑制了朝着包含生物发生的凝聚模型的运动。采用原子力显微镜，我们确定有三层结构，一个外膜是膜双层的厚度，中间网络层和 下面的晶体层状层。遗传研究表明，中间网络至少部分由PHAP组成，并且PHAP可能会易位到周期。因此，似乎通过 将蛋白质和/或蛋白质移动到周期质，并通过细胞质膜萌芽到细胞质中，从而促进了细胞质膜作为包膜的获得。这项研究的目的是证明或反驳这一假设。研究的具体目的是：1）明确证明周期 通过荧光定位和亚细胞级分的蛋白质印迹分析的定位，2）表明，包含包膜是通过蛋白质组学分析从细胞质膜得出的，以及3）表征蛋白质，表征蛋白质，这些蛋白质是瞬时和永久性结合至PHAP的蛋白质，希望能够阐明包含生物的机制。最终，该研究的目的是扩大我们的包容性生物发生知识，以至于该过程可以用于医疗应用。例如，可以预见的是，可以插入生物活性化合物，而不是将聚合物插入包含中，从而将其纳入吸毒工具。