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    DERIVES DE L'INOSITOL-PHOSPHATE ET PROCEDE DE DETECTION DE L'INOSITOL-1-PHOSPHATE
    2.
    发明公开
    DERIVES DE L'INOSITOL-PHOSPHATE ET PROCEDE DE DETECTION DE L'INOSITOL-1-PHOSPHATE 有权
    INOSITPHOSPHATDERIVATE和检测方法的肌醇-1-磷酸

    公开(公告)号：EP1817319A2

    公开(公告)日：2007-08-15

    申请号：EP05824048.2

    申请日：2005-12-02

    申请人： CIS BIO INTERNATIONAL

    发明人： BAZIN, Hervé ,  ANSANAY, Hervé ,  TRINQUET, Eric ,  MATHIS, Gérard

    IPC分类号： C07F9/17

    CPC分类号： C07F9/6561 ,  C07F9/117 ,  C07F9/5721 ,  C07F9/65517 ,  C07F9/65583

    摘要： The invention relates to inositol-phosphate derivatives, in which the inositol-phosphate is substituted by one or two reactive groups G or one or two conjugated molecules M or substances, said reactive group(s) G or said substance(s) or molecule(s) M being linked to IPI via a linkage group L. According to the invention, M is selected from the following group: a tracer, an immunogen, a member of a pair of binding partners, a solid support. The invention is suitable for tools that are used to study the cycle of inositols-phosphates and thus, indirectly, to study seven-domain transmembrane receptors that are coupled to the phospholipase C, receptors having a tyrosine-kinase activity and, generally, enzymes involved in variations in IPI intracellular concentration.

    METHODE DE DETECTION DE L'INTERNALISATION DE PROTEINES MEMBRANAIRES
    4.
    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。

    METHOD FOR THE DETECTION OF POST-TRANSLATIONAL MODIFICATIONS
    5.
    发明公开
    METHOD FOR THE DETECTION OF POST-TRANSLATIONAL MODIFICATIONS 有权
    检测方法的翻译后修饰

    公开(公告)号：EP2087355A2

    公开(公告)日：2009-08-12

    申请号：EP07859356.3

    申请日：2007-11-27

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

    发明人： TRINQUET, Eric ,  BRINKER, Achim ,  CLARET, Emmanuel ,  MATHIS, Gérard

    IPC分类号： G01N33/574

    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.