Mechanisms of Clathrin-Independent Endocytosis

网格蛋白独立的内吞作用机制

• 批准号: 8746686
• 负责人: Julie G Donaldson
• 金额: $ 59.26万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8746686
• 关键词: Biochemical; Biological; Biological Assay; CD44 gene; Cell Surface Proteins; Cells; Clathrin; Communication; Cytoplasmic Tail; Destinations; Endocytosis; Extracellular Matrix; Goals; Integrins; Matrix Metalloproteinases; Mediating; Molecular; Molecular Genetics; Nutrient; Proteins; Recycling; S-nitro-N-acetylpenicillamine; Signal Transduction; Sorting - Cell Movement; Technology; Travel; inhibitor/antagonist; interest; trafficking

We have been studying clathrin-independent forms of endocytosis (CIE) and have identified a number of endogenous PM proteins that enter cells through this mechanism. We have begun to study these proteins in detail in an attempt to understand how they travel in cells and whether they specifically interact with cellular machinery. We have identified signals in the cytoplasmic tails of CD44, CD98 and CD147 that are responsible for their altered trafficking and are looking for cellular machinery that is responsible for recognition and sorting of these signals. Understanding how these proteins move into and out of cells is important because these proteins are involved in interaction with the extracellular matrix (CD44), are involved in nutrient transport (CD98) and interact with integrins and matrix metalloproteinases (CD147). To facilitate these studies we are using SNAP-tag technology to quantify endocytosis and recycling of specific cell surface proteins. We are currently looking for compounds that inhibit or stimulate CIE. We have also been interested in examining the communication between CIE and clathrin-mediated endocytosis (CME). In examining the effects of Pitstop, a newly discovered inhibitor of CME, we found that it was also a potent inhibitor of CIE. Pitstop had been identified through a biochemical assay and although it does inhibit CME, it has broader effects.

我们一直在研究网状蛋白非依赖性内吞作用(CIE)，并鉴定了一些通过这一机制进入细胞的内源性PM蛋白。我们已经开始详细研究这些蛋白质，试图了解它们是如何在细胞中传播的，以及它们是否与细胞机械特定地相互作用。我们已经在CD44、CD98和CD147的细胞质尾巴中发现了负责它们改变运输的信号，并正在寻找负责识别和分类这些信号的细胞机制。了解这些蛋白质如何进出细胞是很重要的，因为这些蛋白质参与与细胞外基质(CD44)的相互作用，参与营养物质的运输(CD98)，并与整合素和基质金属蛋白酶(CD147)相互作用。 为了促进这些研究，我们正在使用SNAP-Tag技术来量化特定细胞表面蛋白的内吞和循环。我们目前正在寻找抑制或刺激CIE的化合物。我们也有兴趣研究CIE和笼蛋白介导的内吞作用(CME)之间的联系。在检测新发现的CME抑制剂PITSTOP的效果时，我们发现它也是一种有效的CIE抑制剂。Pit Stop是通过生化分析鉴定出来的，虽然它确实能抑制CME，但它有更广泛的影响。