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    Heterologous G protein coupled receptors expressed in yeast, their fusion with G proteins and use thereof in bioassay
    1.
    发明申请
    Heterologous G protein coupled receptors expressed in yeast, their fusion with G proteins and use thereof in bioassay 审中-公开
    在酵母中表达的异源G蛋白偶联受体，它们与G蛋白的融合以及其在生物测定中的用途

    公开(公告)号：US20090081764A1

    公开(公告)日：2009-03-26

    申请号：US11637196

    申请日：2006-12-12

    申请人： Mark Henry Pausch ,  Bradley Alton Ozenberger ,  John Richard Hadcock ,  Laura Alicia Price ,  Eileen Marie Kajkowski ,  Donald Richard Kirsch ,  Deborah Tardy Chaleff

    发明人： Mark Henry Pausch ,  Bradley Alton Ozenberger ,  John Richard Hadcock ,  Laura Alicia Price ,  Eileen Marie Kajkowski ,  Donald Richard Kirsch ,  Deborah Tardy Chaleff

    IPC分类号： C12N1/19

    CPC分类号： C07K14/723 ,  C07K2319/00

    摘要： The present invention is directed to expression vectors and yeast cells transformed therewith containing a first heterologous nucleotide sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second nucleotide sequence which codes for all or a portion of a G protein αβγ complex. Said heterologous protein is physically expressed in a host cell membrane in proper orientation for both stereoselective binding of ligands, as well as functional interaction with G proteins on the cytoplasmic side of the cell membrane. In some embodiments, a nucleotide sequence encoding a heterologous or chimeric Gα protein is expressed in conjunction with nucleotide sequences from the yeast G protein βγ subunits. A second aspect of the present invention provides expression vectors and yeast cells transformed therewith encoding chimeric yeast/heterologous G protein coupled receptors. A third aspect of the present invention is directed to methods of assaying compounds using such expression constructs and yeast cell expression systems to determine the effects of ligand binding to the heterologous receptors expressed in the systems.

    摘要翻译： 本发明涉及转化的表达载体和酵母细胞，其含有编码G蛋白偶联受体（例如生长抑素受体）的第一异源核苷酸序列，以及编码全部或一部分 G蛋白质字母复合体。 所述异源蛋白质在适当取向的宿主细胞膜中物理表达，用于配体的立体选择性结合，以及与细胞膜细胞质侧上的G蛋白的功能相互作用。 在一些实施方案中，编码异源或嵌合Galpha蛋白的核苷酸序列与来自酵母G蛋白betagamma亚基的核苷酸序列一起表达。 本发明的第二方面提供了用其编码嵌合酵母/异源G蛋白偶联受体的转化的表达载体和酵母细胞。 本发明的第三方面涉及使用这种表达构建体和酵母细胞表达系统测定化合物的方法，以确定配体与体系中表达的异源受体结合的作用。

    Method for detecting ligand binding to G protein coupled receptors
    8.
    发明授权
    Method for detecting ligand binding to G protein coupled receptors 失效
    检测配体结合G蛋白偶联受体的方法

    公开(公告)号：US06406871B1

    公开(公告)日：2002-06-18

    申请号：US08696924

    申请日：1996-10-15

    申请人： Mark Henry Pausch ,  Bradley Alton Ozenberger ,  John Richard Hadcock ,  Laura Alicia Price ,  Eileen Marie Kajkowski ,  Donald Richard Kirsch ,  Deborah Tardy Chaleff

    发明人： Mark Henry Pausch ,  Bradley Alton Ozenberger ,  John Richard Hadcock ,  Laura Alicia Price ,  Eileen Marie Kajkowski ,  Donald Richard Kirsch ,  Deborah Tardy Chaleff

    IPC分类号： G01N33569

    CPC分类号： C07K14/723 ,  C07K2319/00

    摘要： The present invention is directed to expression vectors and yeast cells transformed therewith containing a first heterologous nucleotide sequence which codes for a G protein-coupled receptor, for example, the somatostatin receptor, and a second nucleotide sequence which codes for all or a portion of a G protein &agr;&bgr;&ggr; complex. Said heterologous protein is physically expressed in a host cell membrane in proper orientation for both stereoselective binding of ligands, as well as functional interaction with G proteins on a cytoplasmic side of the cell membrane. In some embodiments, a nucleotide sequence encoding a heterologous or chimeric G&agr; protein is expressed in conjunction with nucleotide sequences from the yeast G protein &bgr;&ggr; subunits. A second aspect of the present invention provides expression vectors and yeast cells transformed therewith encoding chimeric yeast/heterologous G protein coupled receptors. A third aspect of the present invention is directed to methods of assaying compounds using such expression constructs and yeast cell expression systems to determine the effects of ligand binding to the heterologous receptors expressed in the systems.

    摘要翻译： 本发明涉及用其转化的表达载体和酵母细胞，其含有编码G蛋白偶联受体（例如生长抑素受体）的第一异源核苷酸序列，以及编码全部或一部分 G蛋白质字母复合体。 所述异源蛋白质在适当取向的宿主细胞膜中物理表达，用于配体的立体选择性结合，以及与细胞膜细胞质侧上的G蛋白的功能相互作用。 在一些实施方案中，编码异源或嵌合Galpha蛋白的核苷酸序列与来自酵母G蛋白betagamma亚基的核苷酸序列一起表达。 本发明的第二方面提供了用其编码嵌合酵母/异源G蛋白偶联受体的转化的表达载体和酵母细胞。 本发明的第三方面涉及使用这种表达构建体和酵母细胞表达系统测定化合物的方法，以确定配体与体系中表达的异源受体结合的作用。