Optical Investigations of the Mast Cell MAFA Regulatory Protein

肥大细胞 MAFA 调节蛋白的光学研究

• 批准号: 9807822
• 负责人: B.George Barisas
• 金额: $ 40万
• 依托单位: Colorado State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9807822&HistoricalAwards=false
• 关键词: Optical; Investigations; Mast; Cell; MAFA

9807822 BARISAS The long-term goal of this project is to understand in biophysical terms the mechanism of function of the mast cell function-associated antigen(MAFA) on rat 2H3-RBL cells. MAFA is a type II membrane protein with a C-type lectin extracellular domain and a cytoplasmic YSTL motif. Crosslinking of this protein inhibits signaling by the type I Fce receptor (FceR1) despite the presence of only one MAFA per twenty FceR1. The interactions of MAFA with FceR1 and other membrane proteins will be examined through optical approaches including time-resolved phosphorescence anisotropy, fluorescence photobleaching recovery and fluorescence energy transfer. Differences in these interactions among cells receiving various stimulatory and inhibitory stimuli, such as clustering FceR1 by antigen or clustering MAFA by the MAFA-specific antibody G63 will be examined. Finally, this overall experimental approach will be extended to chimeric molecules derived from MAFA and expressed in cells capable of FceR1 signaling. This project provides an excellent opportunity to examine how cell surface molecules of one kind can regulate the function of another molecule present in much larger numbers. The use of molecular biological methods to produce selectively-modified receptors and signaling molecules in combination with biophysical approaches to measuring quantitative parameters of these molecules on living cells provides a powerful approach by which to address this problem. It also provides an opportunity to refine, for use by the larger biophysical community, laser optical techniques for examining the behavior of cell membrane proteins. These methods are a strength of this laboratory.

小行星9807822 本项目的长期目标是从生物物理学角度了解肥大细胞功能相关抗原（MAFA）对大鼠2 H3-RBL细胞的作用机制。 MAFA是具有C型凝集素胞外结构域和胞质YSTL基序的II型膜蛋白。 这种蛋白质的交联抑制I型Fce受体（FceRl）的信号传导，尽管每20个FceRl仅存在一个MAFA。 MAFA与FceR 1和其他膜蛋白的相互作用将通过光学方法，包括时间分辨磷光各向异性，荧光光漂白恢复和荧光能量转移检查。 将检查接受各种刺激性和抑制性刺激的细胞之间这些相互作用的差异，例如通过抗原聚集FceR 1或通过MAFA特异性抗体G63聚集MAFA。 最后，这一整体实验方法将扩展到嵌合分子衍生自MAFA和表达的细胞能够FceR 1信号。 这个项目提供了一个很好的机会来研究一种细胞表面分子如何调节大量存在的另一种分子的功能。 使用分子生物学方法来产生选择性修饰的受体和信号分子，结合生物物理方法来测量活细胞上这些分子的定量参数，提供了解决这个问题的有力方法。 它还提供了一个机会，以完善，由更大的生物物理界使用，激光光学技术检查细胞膜蛋白的行为。 这些方法是本实验室的强项。