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    PROCEDE DE DETECTION DE COMPOSES MODULATEURS DE DIMERES DE PROTEINES MEMBRANAIRES A DOMAINE VFT
    2.
    发明公开
    PROCEDE DE DETECTION DE COMPOSES MODULATEURS DE DIMERES DE PROTEINES MEMBRANAIRES A DOMAINE VFT 有权
    检测方法的VFT域膜蛋白调节性化合物的二聚体

    公开(公告)号：EP2425249A1

    公开(公告)日：2012-03-07

    申请号：EP10727062.1

    申请日：2010-04-29

    申请人： CIS Bio International ,  Centre National de la Recherche Scientifique

    发明人： DOUMAZANE, Etienne ,  ZWIER, Jurriaan ,  TRINQUET, Eric ,  PIN, Jean-Philippe

    IPC分类号： G01N33/542

    CPC分类号： G01N33/542 ,  G01N33/9426 ,  G01N2333/726 ,  G01N2500/04

    摘要： The invention relates to a method for selecting compounds having a modulating effect on the activation state of a dimer of VFT domain proteins expressed in cellular membranes in a measuring medium, said dimer including a first protein and a second protein which are identical or different, wherein the method includes the following steps: (a) marking the first and second proteins in the N-terminal portion of the VFT domain by members of a FRET partner pair, the Förster radius (R
    0 ) of said pair ranging between 20 and 55 Å; (b) measuring the FRET signal in the absence and in the presence of the compound to be tested within a predetermined time window; (c) selecting the compound to be tested as a modulating compound if a difference in the FRET signal in the absence and in the presence of the compound to be tested is measured during step (b). The invention can be used in the research for new drugs and new taste modulators.

    METHODE DE DETECTION DE L'INTERNALISATION DE PROTEINES MEMBRANAIRES
    9.
    发明公开
    METHODE DE DETECTION DE L'INTERNALISATION DE PROTEINES MEMBRANAIRES 有权
    方法检测蛋白质内膜

    公开(公告)号：EP2329272A2

    公开(公告)日：2011-06-08

    申请号：EP09740420.6

    申请日：2009-07-30

    申请人： Cis-Bio International

    发明人： ZWIER, Jurriaan ,  POOLE, Robert ,  ANSANAY, Hervé ,  FINK, Michel ,  TRINQUET, Eric

    IPC分类号： G01N33/566 ,  G01N33/58

    CPC分类号： G01N33/542 ,  G01N33/5035 ,  G01N33/58 ,  G01N2458/40 ,  Y10T436/13

    摘要： A method for detecting the internalization of a transmembrane protein of interest expressed at the surface of a cell, comprising the following steps: a) labelling the protein of interest with a fluorescent metal complex, the lifetime of which is greater than 0.1 ms; b) adding to the reaction medium a composition capable of causing the internalization of the protein of interest; c) adding to the reaction medium a modulating agent selected from: a. a fluorescent or nonfluorescent FRET acceptor compound compatible with said fluorescent metal complex, the final concentration of which in the reaction medium is greater than 10
    ‑7 M; b. a reducing agent, the redox potential of which is less than +0.1 V and preferably between 0.25 and 0.75 V; c. an agent which binds specifically, by noncovalent bonding, with the fluorescent metal complex; d. a metal ion which competes with the rare earth so as to form a nonfluorescent metal complex; d) measuring the luminescence emitted by the reaction medium at the emission wavelength of the fluorescent metal complex and/or at the emission wavelength of the modulating compound when said compound is a fluorescent acceptor compound; e) comparing the signal measured in step d) with a reference signal measured on cells having been subjected only to steps a) and c). Use: Method for detecting membrane protein internalization.

    摘要翻译： 一种检测细胞表面表达的感兴趣的跨膜蛋白的内化的方法，包括以下步骤：a）用荧光金属配合物标记感兴趣的蛋白质，其寿命大于0.1ms; b）向反应介质中加入能够引起感兴趣蛋白质内化的组合物; c）向反应介质中加入选自以下的调节剂：a。 与所述荧光金属配合物相容的荧光或非荧光FRET受体化合物，其在反应介质中的最终浓度大于10 -7 M; 湾 还原剂，其氧化还原电位低于+ 0.1V，优选在0.25和0.75V之间; C。 通过非共价键特异性结合荧光金属配合物的试剂; d。 与稀土竞争以形成无荧光金属络合物的金属离子; d）当所述化合物是荧光受体化合物时，在荧光金属络合物的发射波长处和/或在调制化合物的发射波长处测量由反应介质发射的发光; e）将步骤d）中测得的信号与仅在经过步骤a）和c）的细胞上测量的参考信号进行比较。 用途：检测膜蛋白内在化的方法。