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    • 3. 发明授权
    • Method for assaying for a substance that affects an SH2 phosphorylated
ligand regulatory system
    • 测定影响SH2磷酸化配体调节系统的物质的方法
    • US5667980A
    • 1997-09-16
    • US221699
    • 1994-03-28
    • Anthony PawsonMichael Reedijk
    • Anthony PawsonMichael Reedijk
    • G01N33/566G01N33/573G01N33/74C12Q1/48G01N33/53G01N33/68
    • G01N33/573G01N33/566G01N33/74G01N2333/71G01N2333/9121G01N2500/20
    • A method for assaying a medium for the presence of a substance that affects an SH2-phosphorylated ligand regulatory system. The method employs an SH2-like domain or a subdomain thereof and a phosphorylated ligand. The phosphophorylated ligand is capable of interacting with the SH2-like domain or a subdomain thereof to form an SH2-phosphorylated ligand complex. The SH2-like domain or subdomain and/or the phosphorylated ligand are present in a known concentration. The SH2-like domain or a subdomain thereof and the phosphorylated ligand are incubated with a substance which is suspected of affecting an SH2-phosphorylated ligand regulatory system. The method is carried out under conditions which permit the formation of the SH2-phosphorylated ligand complex. SH2-phosphorylated ligand complex, free SH2-like domain or subdomains thereof, or non-complexed phosphorylated ligand are assayed. The invention also relates to an isolated SH2-phosphorylated ligand complex; a method of using an isolated SH2-like domain or a subdomain thereof to screen for phosphorylated ligands which are active in an SH2-phosphorylated ligand regulatory system; a method of using an isolated SH2-like domain or a subdomain thereof to regulate the interaction of a signalling protein with a related phosphorylated ligand; and a pharmaceutical composition comprising an isolated SH2-like domain or a subdomain thereof for use as an agohist or antagonist of the interaction of a signalling protein with a related phosphorylated ligand.
    • 用于测定培养基中存在影响SH2磷酸化配体调节系统的物质的方法。 该方法采用SH2样结构域或其亚结构域和磷酸化配体。 磷酰化配体能够与SH2样结构域或其亚结构域相互作用形成SH2-磷酸化的配体复合物。 SH2样结构域或亚结构域和/或磷酸化配体以已知浓度存在。 将SH2样结构域或其亚结构域和磷酸化配体与怀疑影响SH2-磷酸化配体调节系统的物质一起温育。 该方法在允许形成SH2-磷酸化配体络合物的条件下进行。 测定SH2-磷酸化配体复合物,游离SH2样结构域或其子结构域,或非络合磷酸化配体。 本发明还涉及分离的SH2-磷酸化配体复合物; 使用分离的SH2样结构域或其亚结构域筛选在SH2磷酸化配体调节系统中有活性的磷酸化配体的方法; 使用分离的SH2样结构域或其亚结构域来调节信号蛋白与相关磷酸化配体的相互作用的方法; 以及包含分离的SH2样结构域或其子结构域的药物组合物,用作信号蛋白与相关磷酸化配体的相互作用的前组或拮抗剂。
    • 4. 发明授权
    • Method for measuring glycosyltransferase activity
    • 测定糖基转移酶活性的方法
    • US5501957A
    • 1996-03-26
    • US293940
    • 1994-08-22
    • James W. DennisKatherine A. SiminovitchAlessandro Datti
    • James W. DennisKatherine A. SiminovitchAlessandro Datti
    • C12Q1/48A01N43/04C12Q1/00G01N33/00
    • C12Q1/48Y10S435/81Y10T436/141111Y10T436/143333
    • A method of assaying for glycosyltransferase activity in a sample. In a first step, a sample is reacted with a first sugar donor and an acceptor substrate to produce a transferase product. The first sugar donor and acceptor substrate are selected such that the sugar from the first sugar donor is capable of being transferred to the acceptor substrate in the presence of the glycosyltransferase to be assayed. In a second step, the transferase product is reacted with a second sugar donor having a sugar which is labelled with a labelling agent and an enzyme which is capable of transferring the sugar from the second sugar donor to the transferase product to produce a labelled transferase product and which has a higher affinity for the glycosyltransferase product compared to the affinity of the glycosyltransferase for the acceptor substrate. The labelling agent activity of the labelled transferase product or unreacted second sugar donor is assayed to determine transferase activity in the sample. A kit for assaying for glycosyltransferase activity in a sample is also described.
    • 测定样品中糖基转移酶活性的方法。 在第一步中,将样品与第一糖供体和受体底物反应以产生转移酶产物。 选择第一糖供体和受体底物使得来自第一糖供体的糖能够在待测定的糖基转移酶存在下转移到受体底物。 在第二步中,将转移酶产物与具有用标记试剂标记的糖的第二糖供体进行反应,所述糖能够将糖从第二糖供体转移到转移酶产物以产生标记的转移酶产物 并且与糖基转移酶对受体底物的亲和力相比,其对糖基转移酶产物具有更高的亲和力。 测定标记的转移酶产物或未反应的第二糖供体的标记试剂活性,以确定样品中的转移酶活性。 还描述了用于测定样品中糖基转移酶活性的试剂盒。