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Targeted fusion proteins and methods for the characterization of cellular membrane domains

机译:靶向融合蛋白和表征细胞膜结构域的方法

摘要

Cell membranes containing glycolipid-enriched membrane (GEM) and non-glycolipid-enriched membrane (non-GEM) domains are targeted using fusion proteins that are anchored in the cell membrane. Fusion proteins to target GEM (or non-GEM) domains are comprised of a selected fluorescent polypeptide, a membrane-targeting sequence of p56Lck (or pp60c-Src for non-GEM domains) and a linker inserted between the polypeptide and the membrane targeting sequence. Localization of fusion proteins in GEM and non-GEM domains is assessed using techniques including confocal microscopy, fluorescence-based techniques, and membrane fractionation. Using these techniques, compounds are screened for their effect on GEM and non-GEM domains of live cells. These fusion proteins therefore represent useful tools for studying subcellular trafficking and the function of discrete compartments in the plasma membrane.
机译:使用锚定在细胞膜中的融合蛋白靶向含有糖脂富集膜(GEM)和非糖脂富集膜(non-GEM)域的细胞膜。靶向GEM(或非GEM)结构域的融合蛋白由选定的荧光多肽,p56 Lck (或pp60 c-Src )的膜靶向序列组成-GEM结构域)和插入多肽和膜靶向序列之间的接头。融合蛋白在GEM和非GEM域中的定位使用共聚焦显微镜,基于荧光的技术和膜分离技术进行评估。使用这些技术,筛选化合物对活细胞的GEM和非GEM域的影响。因此,这些融合蛋白代表了研究亚细胞运输和质膜中离散隔室功能的有用工具。

著录项

  • 公开/公告号US6838263B2

    专利类型

  • 公开/公告日2005-01-04

    原文格式PDF

  • 申请/专利权人 WILLIAM RODGERS;

    申请/专利号US20020191767

  • 发明设计人 WILLIAM RODGERS;

    申请日2002-07-09

  • 分类号C12P2106;

  • 国家 US

  • 入库时间 2022-08-21 22:19:02

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