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1. 发明授权

US08697380B2 Method for detecting compounds modulating dimers of VFT domain membrane proteins 有权
标题翻译：检测调节VFT结构域膜蛋白二聚体的化合物的方法
公开(公告)号：US08697380B2
公开(公告)日：2014-04-15
申请号：US13266222
申请日：2010-04-29
申请人： Etienne Doumazane , Jurriaan Zwier , Eric Trinquet , Jean-Phillippe Pin
发明人： Etienne Doumazane , Jurriaan Zwier , Eric Trinquet , Jean-Phillippe Pin
IPC分类号： C12Q1/02 , C12Q1/34 , C12Q1/48
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 cell membranes present in a measuring medium, said dimer consisting of a first protein and of a second protein, said proteins being identical or different, wherein this method comprises the following steps: (a) labeling the first and second proteins in the N-terminal portion of their VFT domains with the members of a pair of FRET partners, the Förster radius (R0) of said pair being between 20 and 55 Å; (b) measuring the FRET signal in the absence and in the presence of the test compound within a predetermined time window; (c) selecting the test compound as a modulating compound if a difference in FRET signal in the absence and in the presence of test compound is measured in step (b). The invention can be used in the search for new medicaments and new taste modulators.
摘要翻译：本发明涉及一种选择对存在于测量介质中的细胞膜中表达的VFT结构域蛋白的二聚体的活化状态具有调节作用的化合物的方法，所述二聚体由第一种蛋白质和第二种蛋白质组成，所述蛋白质相同或不同，其中该方法包括以下步骤：（a）用一对FRET伴侣的成员将其VFT结构域的N末端部分中的第一和第二蛋白质标记，所述第一和第二蛋白质的所述F 对在20和55之间; （b）在预定时间窗内测试化合物不存在和存在时测量FRET信号; （c）如果在步骤（b）中测定了在不存在和存在测试化合物的情况下FRET信号的差异，则选择测试化合物作为调节化合物。本发明可用于寻找新的药物和新的调味剂。

2. 发明授权

US08663928B2 Method for the detection of post-translational modifications 有权
标题翻译：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。

3. 发明授权

US09024022B2 Substrates of O6-alkylguanine-DNA alkyltransferase and mutants thereof 有权
标题翻译： O6-烷基鸟嘌呤-DNA烷基转移酶的底物及其突变体
公开(公告)号：US09024022B2
公开(公告)日：2015-05-05
申请号：US13120197
申请日：2009-09-22
申请人： Emmanuel Bourrier , Michel Laget , Laurent Lamarque , Norbert Tinel , Eric Trinquet , Hervé Bazin
发明人： Emmanuel Bourrier , Michel Laget , Laurent Lamarque , Norbert Tinel , Eric Trinquet , Hervé Bazin
IPC分类号： C07D473/18 , C07D239/47 , C07D403/14 , C07D487/18 , C07D519/00
CPC分类号： C07D239/47 , C07D403/14 , C07D473/18 , C07D487/18 , C07D519/00
摘要： The invention relates to compounds of formula (I′): in which A, L2, M and B are as defined in the description. These compounds are substrates of O6-alkylguanine-DNA alkyltransferase and mutants thereof.
摘要翻译：本发明涉及式（I'）化合物：其中A，L2，M和B如说明书中所定义。这些化合物是O6-烷基鸟嘌呤-DNA烷基转移酶及其突变体的底物。

4. 发明申请

US20120190048A1 METHOD FOR DETERMINING THE BINDING OF A GIVEN COMPOUND TO A MEMBRANE RECEPTOR 有权
标题翻译：用于确定化合物与膜受体结合的方法
公开(公告)号：US20120190048A1
公开(公告)日：2012-07-26
申请号：US13388150
申请日：2010-08-12
申请人： Eric Trinquet , Jurriaan Zwier , Gérard Mathis , Jean-Philippe Pin , Thierry Durroux
发明人： Eric Trinquet , Jurriaan Zwier , Gérard Mathis , Jean-Philippe Pin , Thierry Durroux
IPC分类号： G01N33/566
CPC分类号： G01N33/542 , G01N33/74
摘要： The invention relates to a method for determining whether a test compound binds preferentially to a membrane receptor R1 or to a membrane receptor R2, these receptors being known to be expressed on the surface of the cells in monomeric, homodimeric or heterodimeric form. This method is applied using one or two FRET partner pairs.
摘要翻译：本发明涉及一种用于确定测试化合物优先与膜受体R1或膜受体R2结合的方法，已知这些受体以单体，同二聚体或异二聚体形式在细胞表面表达。使用一个或两个FRET伙伴对应用此方法。

5. 发明授权

US08178704B2 Inositol-phosphate derivatives and method of detecting inositol-1-phosphate 有权
标题翻译：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。

6. 发明授权

US08999653B2 Method for detecting membrane protein internalization 有权
标题翻译：膜蛋白内化检测方法
公开(公告)号：US08999653B2
公开(公告)日：2015-04-07
申请号：US13056738
申请日：2009-07-30
申请人： Jurriaan Zwier , Robert Poole , Herve Ansanay , Michel Fink , Eric Trinquet
发明人： Jurriaan Zwier , Robert Poole , Herve Ansanay , Michel Fink , Eric Trinquet
IPC分类号： G01N33/53 , C12Q1/00 , G01N21/76 , G01N33/542 , G01N33/50 , G01N33/58
CPC分类号： G01N33/542 , G01N33/5035 , G01N33/58 , G01N2458/40 , Y10T436/13
摘要： The instant invention provides for methods for detecting the internalization of a transmembrane protein of interest expressed at the surface of a cell. More specifically, the methods involve (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 (1) a modulating agent which is a fluorescent FRET acceptor compound compatible with the fluorescent metal complex, the final concentration of which in the reaction medium is greater than 10−7M; or (2) a reducing agent, the redox potential of which is less than +0.1 V and preferably between 0.25 and 0.75 V; or (3) an agent which binds specifically, by non-covalent bonding, with the fluorescent metal complex; (d) adding a metal ion which competes with the rare earth so as to form a non-fluorescent 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 the compound is a fluorescent acceptor compound; and (e) comparing the signal measured in step d) with a reference signal measured on cells having been subjected only to steps a) and c).
摘要翻译：本发明提供了用于检测在细胞表面表达的感兴趣的跨膜蛋白的内在化的方法。更具体地说，所述方法包括（a）用荧光金属络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的蛋白质内化的组合物，（c）向反应介质（1）中加入调节剂，其是与荧光金属络合物相容的荧光FRET受体化合物，其最终浓度在反应介质中的浓度大于10-7M; 或（2）还原剂，其氧化还原电位小于+ 0.1V，优选0.25-0.75V; 或（3）通过非共价键合与荧光金属络合物特异性结合的试剂; （d）加入与稀土竞争的金属离子以形成非荧光金属络合物; （d）当化合物是荧光受体化合物时，测量在荧光金属络合物的发射波长处和/或调节化合物的发射波长处的反应介质发射的发光; 和（e）将步骤d）中测量的信号与在仅经受步骤a）和c）的细胞上测量的参考信号进行比较。

7. 发明申请

US20120009599A1 METHOD FOR DETECTING MEMBRANE PROTEIN INTERNALIZATION 有权
标题翻译：检测膜蛋白内含物的方法
公开(公告)号：US20120009599A1
公开(公告)日：2012-01-12
申请号：US13056738
申请日：2009-07-30
申请人： Jurriaan Zwier , Robert Poole , Herve Ansanay , Michel Fink , Eric Trinquet
发明人： Jurriaan Zwier , Robert Poole , Herve Ansanay , Michel Fink , Eric Trinquet
IPC分类号： G01N21/64
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 10−7M; 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).
摘要翻译：一种用于检测在细胞表面表达的目标跨膜蛋白的内在化的方法，包括以下步骤：a）用荧光金属络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目的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。与所述荧光金属络合物相容的荧光或非荧光FRET受体化合物，其最终浓度在反应介质中大于10-7M; b。还原剂，其氧化还原电位小于+0.1V，优选0.25至0.75V; C。通过非共价键合与荧光金属络合物特异性结合的试剂; d。与稀土竞争以形成非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，测量在荧光金属络合物的发射波长处和/或调节化合物的发射波长处的反应介质发射的发光; e）将步骤d）中测量的信号与在仅经受步骤a）和c）的细胞上测量的参考信号进行比较。

8. 发明申请

US20110236952A1 NOVEL SUBSTRATES OF O6-ALKYLGUANINE-DNA ALKYLTRANSFERASE AND MUTANTS THEREOF 有权
标题翻译： O6-谷氨酰胺-DNA烷基转移酶及其突变体的新基质
公开(公告)号：US20110236952A1
公开(公告)日：2011-09-29
申请号：US13120197
申请日：2009-09-22
申请人： Emmanuel Bourrier , Michel Laget , Laurent Lamarque , Norbet Tinel , Eric Trinquet , Hervé Bazin
发明人： Emmanuel Bourrier , Michel Laget , Laurent Lamarque , Norbet Tinel , Eric Trinquet , Hervé Bazin
IPC分类号： C12N9/96 , C07D473/18 , C07D403/14 , C07F5/00 , C07K14/77 , C07K14/36 , C07K2/00
CPC分类号： C07D239/47 , C07D403/14 , C07D473/18 , C07D487/18 , C07D519/00
摘要： The invention relates to compounds of formula (I′): in which A, L2, M and B are as defined in the description.These compounds are substrates of O6-alkylguanine-DNA alkyltransferase and mutants
摘要翻译：本发明涉及式（I'）化合物：其中A，L2，M和B如说明书中所定义。这些化合物是O6-烷基鸟嘌呤-DNA烷基转移酶和突变体的底物

9. 发明授权

US07872243B2 Method for improving the detection of fluorescence signals during a resonance energy transfer 有权
标题翻译：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。

10. 发明申请

US20090220988A1 METHOD OF REVEALING A BIOLOGICAL PROCESS USING A FRET MEASUREMENT 有权
标题翻译：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。

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