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    Method for the detection of post-translational modifications
    3.
    发明授权
    Method for the detection of post-translational modifications 有权
    检测翻译后修饰的方法

    公开(公告)号：US08663928B2

    公开(公告)日：2014-03-04

    申请号：US12516333

    申请日：2007-11-27

    申请人： Eric Trinquet ,  Achim Brinker ,  Emmanuel Claret ,  Gérard Mathis

    发明人： Eric Trinquet ,  Achim Brinker ,  Emmanuel Claret ,  Gérard Mathis

    IPC分类号： G01N33/68 ,  G01N33/53 ,  G01N33/58

    CPC分类号： G01N33/542 ,  C12Q1/37 ,  C12Q1/48 ,  C12Q1/485

    摘要： Method for the detection in homogeneous medium of a post-translational modification of a proteinaceous substrate catalyzed by a cell enzyme, characterized in that the post-translational modification reaction takes place in intact living cells, in that these cells comprise a heterologous expression vector coding for a fusion protein comprising the proteinaceous substrate and a first coupling domain and in that it comprises the following stages: (i) Incubation of the cells in the presence or in the absence of a compound to be tested capable of modulating the activity of said enzyme, (ii) Addition to the reaction medium of a first fluorescent compound member of a first pair of FRET partners covalently bonded to a coupling agent capable of binding specifically to the first coupling domain present on the proteinaceous substrate, (iii) Addition to the reaction medium of a second fluorescent compound member of this first pair of FRET partners, and covalently bonded to a binding domain specific to the site of the proteinaceous substrate having undergone the post-translational modification and not binding to the non-modified proteinaceous substrate, (i) Measurement of the FRET signal emitted by the sample, this signal being representative of the quantity of proteinaceous substrate having undergone said post-translational modification; and cells for the implementation of said method.

    摘要翻译： 在均质培养基中检测由细胞酶催化的蛋白质底物的翻译后修饰的方法，其特征在于翻译后修饰反应在完整的活细胞中进行，因为这些细胞包含编码 包含蛋白质底物和第一偶联结构域的融合蛋白，并且其包含以下阶段：（i）在待测化合物存在或不存在下能够调节所述酶的活性的细胞的孵育， （ii）向第一对FRET伴侣的第一荧光化合物成员的反应介质中加入共价键合到能够特异性结合存在于蛋白质底物上的第一偶联结构域的偶联剂，（iii）加入反应介质 的第一对FRET配偶体的第二荧光化合物成员，并共价键合到结合域 c到经过翻译后修饰并且不结合未修饰的蛋白质底物的蛋白质底物的位点，（i）测量由样品发射的FRET信号，该信号代表具有 经过翻译后修改; 以及用于执行所述方法的单元。

    Method for improving the detection of fluorescence signals during a resonance energy transfer
    4.
    发明授权
    Method for improving the detection of fluorescence signals during a resonance energy transfer 有权
    在共振能量转移期间改善荧光信号检测的方法

    公开(公告)号：US07872243B2

    公开(公告)日：2011-01-18

    申请号：US11571331

    申请日：2005-06-27

    申请人： Eric Trinquet ,  Gérard Mathis

    发明人： Eric Trinquet ,  Gérard Mathis

    IPC分类号： G01J1/58

    CPC分类号： G01N21/6428 ,  G01N21/6445

    摘要： The invention concerns the use of the fluorescence polarization phenomenon to improve detection of fluorescent signals during a fluorescence resonance energy transfer (FRET). In particular, the invention concerns a method for improving signal/noise ratio in a FRET measurement. The invention also concerns an apparatus for measuring fluorescence following an energy transfer between a donor fluorescent compound and an acceptor fluorescence compound in a measurement medium.

    摘要翻译： 本发明涉及使用荧光偏振现象来改善在荧光共振能量转移（FRET）期间荧光信号的检测。 特别地，本发明涉及一种在FRET测量中改善信号/噪声比的方法。 本发明还涉及用于在测量介质中的供体荧光化合物和受体荧光化合物之间的能量转移之后测量荧光的装置。

    METHOD OF REVEALING A BIOLOGICAL PROCESS USING A FRET MEASUREMENT
    5.
    发明申请
    METHOD OF REVEALING A BIOLOGICAL PROCESS USING A FRET MEASUREMENT 有权
    使用FRET测量发现生物过程的方法

    公开(公告)号：US20090220988A1

    公开(公告)日：2009-09-03

    申请号：US12065210

    申请日：2006-08-29

    申请人： Eric Trinquet ,  Hervè Ansanay ,  Gérard Mathis

    发明人： Eric Trinquet ,  Hervè Ansanay ,  Gérard Mathis

    IPC分类号： G01N33/53 ,  C12Q1/02

    CPC分类号： G01N33/542 ,  G01N33/6872 ,  Y10S435/968

    摘要： The present invention relates to a method of revealing a biological process using a FRET measurement, which comprises the following steps: incorporating, into a measurement medium containing a lipid membrane, a biological entity X coupled with a first member of a pair of FRET partners and a second biological entity Y coupled with the second member of the pair of FRET partners, the energy-donating member of the pair of FRET partners having a long lifetime and the members of said pair of FRET partners being located on either side of the lipid membrane; exciting the measurement medium at the excitation wavelength of the energy-donating member; and measuring the FRET signal or the variations in said signal emitted in said culture medium.

    摘要翻译： 本发明涉及使用FRET测定揭示生物过程的方法，其包括以下步骤：将含有脂质膜的生物体X与一对FRET伴侣的第一成员结合， 与该对FRET伴侣的第二成员结合的第二生物实体Y，该对FRET伴侣中的能量供给成员具有长寿命，并且所述一对FRET伴侣的成员位于脂质膜的任一侧 ; 在能量供给构件的激发波长下激发测量介质; 以及测量FRET信号或在所述培养基中发射的所述信号的变化。

    Inositol-phosphate derivatives and method of detecting inositol-1-phosphate
    7.
    发明授权
    Inositol-phosphate derivatives and method of detecting inositol-1-phosphate 有权
    肌醇磷酸酯衍生物和肌醇-1-磷酸酯检测方法

    公开(公告)号：US08178704B2

    公开(公告)日：2012-05-15

    申请号：US11792135

    申请日：2005-12-02

    申请人： Hervé Bazin ,  Hervé Ansanay ,  Eric Trinquet ,  Gérard Mathis

    发明人： Hervé Bazin ,  Hervé Ansanay ,  Eric Trinquet ,  Gérard Mathis

    IPC分类号： C07F9/02 ,  G01N33/53

    CPC分类号： C07F9/6561 ,  C07F9/117 ,  C07F9/572 ,  C07F9/65517 ,  C07F9/65583

    摘要： The present invention relates to inositol phosphate derivatives, in which the inositol phosphate is substituted with one or two reactive groups G or one or two conjugated substances or molecules M, said reactive group(s) G or said substance(s) or molecule(s) M being linked to IP1 via a linkage group L, M being chosen from the following group: a tracer, an immunogen, a member of a binding partner pair, a solid support.Application: tools allowing the study of the inositol phosphate cycle and therefore, indirectly, the study of seven transmembrane domain receptors coupled to phospholipase C, receptors having a tyrosine kinase activity, and in general enzymes involved in the variations of the intracellular concentration of IP1.

    摘要翻译： 本发明涉及肌醇磷酸酯衍生物，其中磷酸肌醇被一个或两个反应性基团G或一个或两个共轭物质或分子M取代，所述反应性基团G或所述物质或分子 ）M通过连接基团L连接到IP1，M选自以下组：示踪剂，免疫原，结合配偶体对的成员，固体支持物。 应用：允许研究肌醇磷酸循环的工具，因此间接研究偶联于磷脂酶C的七个跨膜结构域受体，具有酪氨酸激酶活性的受体，以及涉及IP1细胞内浓度变化的一般酶。

    Method of revealing a biological process using a FRET measurement
    9.
    发明授权
    Method of revealing a biological process using a FRET measurement 有权
    使用FRET测量揭示生物过程的方法

    公开(公告)号：US07998694B2

    公开(公告)日：2011-08-16

    申请号：US12065210

    申请日：2006-08-29

    申请人： Eric Trinquet ,  Hervé Ansanay ,  Gérard Mathis

    发明人： Eric Trinquet ,  Hervé Ansanay ,  Gérard Mathis

    IPC分类号： G01N33/567

    CPC分类号： G01N33/542 ,  G01N33/6872 ,  Y10S435/968

    摘要： The present invention relates to a method of revealing a biological process using a FRET measurement, which comprises the following steps: incorporating, into a measurement medium containing a lipid membrane, a biological entity X coupled with a first member of a pair of FRET partners and a second biological entity Y coupled with the second member of the pair of FRET partners, the energy-donating member of the pair of FRET partners having a long lifetime and the members of said pair of FRET partners being located on either side of the lipid membrane; exciting the measurement medium at the excitation wavelength of the energy-donating member; and measuring the FRET signal or the variations in said signal emitted in said culture medium.

    摘要翻译： 本发明涉及使用FRET测定揭示生物过程的方法，其包括以下步骤：将含有脂质膜的生物体X与一对FRET伴侣的第一成员结合， 与该对FRET伴侣的第二成员结合的第二生物实体Y，该对FRET伴侣中的能量供给成员具有长寿命，并且所述一对FRET伴侣的成员位于脂质膜的任一侧 ; 在能量供给构件的激发波长下激发测量介质; 以及测量FRET信号或在所述培养基中发射的所述信号的变化。

    METHOD FOR THE DETECTION OF POST-TRANSLATIONAL MODIFICATIONS
    10.
    发明申请
    METHOD FOR THE DETECTION OF POST-TRANSLATIONAL MODIFICATIONS 有权
    检测翻译后修改的方法

    公开(公告)号：US20100086943A1

    公开(公告)日：2010-04-08

    申请号：US12516333

    申请日：2007-11-27

    申请人： Eric Trinquet ,  Achim Brinker ,  Emmanuel Claret ,  Gérard Mathis

    发明人： Eric Trinquet ,  Achim Brinker ,  Emmanuel Claret ,  Gérard Mathis

    IPC分类号： G01N33/53 ,  C12N5/00 ,  C12N5/071

    CPC分类号： G01N33/542 ,  C12Q1/37 ,  C12Q1/48 ,  C12Q1/485

    摘要： Method for the detection in homogeneous medium of a post-translational modification of a protein substrate catalyzed by a cell enzyme, characterized in that the post-translational modification reaction takes place in intact living cells, in that these cells comprise a heterologous expression vector coding for a fusion protein comprising the protein substrate and a first coupling domain and in that it comprises the following stages: (v) Incubation of the cells in the presence or in the absence of a compound to be tested capable of modulating the activity of said enzyme, (vi) Addition to the reaction medium of a first fluorescent compound member of a first pair of FRET partners covalently bonded to a coupling agent capable of binding specifically to the first coupling domain present on the protein substrate, (vii) Addition to the reaction medium of a second fluorescent compound member of this first pair of FRET partners, and covalently bonded to a binding domain specific to the site of the protein substrate having undergone the post-translational modification and not binding to the non-modified protein substrate, (i) Measurement of the FRET signal emitted by the sample, this signal being representative of the quantity of protein substrate having undergone said post-translational modification; and cells for the implementation of said method.

    摘要翻译： 用于在均匀培养基中检测由细胞酶催化的蛋白质底物的翻译后修饰的方法，其特征在于翻译后修饰反应在完整的活细胞中进行，因为这些细胞包含编码 包含蛋白质底物和第一偶联结构域的融合蛋白，并且其包含以下阶段：（v）在待测化合物存在或不存在下能够调节所述酶的活性的细胞的孵育， （vi）向第一对FRET配偶体的第一荧光化合物成员的反应介质中加入共价结合到能够特异性结合蛋白质底物上存在的第一偶联结构域的偶联剂，（vii）加入到反应介质中 的第一对FRET配偶体的第二荧光化合物成员，并且共价键合到特异于t位点的结合结构域 已经经历了翻译后修饰并且不结合未修饰的蛋白质底物的蛋白质底物，（i）测量由样品发射的FRET信号，该信号代表已经经历所述翻译后蛋白质底物的量 修改; 以及用于执行所述方法的单元。