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    Designer ubiquitin ligases having a non-cleavable SNAP25 domain and E3-ligase domain
    5.
    发明授权
    Designer ubiquitin ligases having a non-cleavable SNAP25 domain and E3-ligase domain 失效
    具有不可切割的SNAP25结构域和E3连接酶结构域的设计者泛素连接酶

    公开(公告)号：US08343743B2

    公开(公告)日：2013-01-01

    申请号：US12482420

    申请日：2009-06-10

    Applicant: George A. Oyler ,  Yien Che Tsai

    Inventor： George A. Oyler ,  Yien Che Tsai

    IPC: C12N9/00 ,  C12N1/20 ,  C12N15/00 ,  C07H21/04 ,  A61K38/43

    CPC classification number: C12N9/93 ,  A61K38/4886 ,  Y02A50/471

    Abstract: The present invention relates to a designer or recombinant ubiquitin ligase molecule that includes a toxin binding domain that is specific for a toxin active fragment, wherein the toxin active fragment is an enzymatically active fragment of one or more toxins or toxin serotypes; and an E3-ligase domain that comprises an E3-ligase or polypeptide that facilitates E2-mediated ubiquitination of the toxin active fragment. In an embodiment, the composition further includes a delivery system that allow the designer ubiquitin ligase to enter the cell. The present invention further includes methods for treating an individual intoxicated with a toxin by administering the designer ubiquitin ligase of the present invention.

    Abstract translation: 本发明涉及一种设计者或重组泛素连接酶分子，其包括对毒素活性片段特异的毒素结合结构域，其中所述毒素活性片段是一种或多种毒素或毒素血清型的酶活性片段; 和E3连接酶结构域，其包含促进E2介导的毒素活性片段的泛素化的E3连接酶或多肽。 在一个实施方案中，组合物还包括允许设计者泛素连接酶进入细胞的递送系统。 本发明还包括通过施用本发明的设计者泛素连接酶来治疗用毒素中毒的个体的方法。

    N-end rule protease activity indication methods and uses thereof
    8.
    发明授权
    N-end rule protease activity indication methods and uses thereof 有权
    N端规则蛋白酶活性指示方法及其用途

    公开(公告)号：US08940482B1

    公开(公告)日：2015-01-27

    申请号：US13159284

    申请日：2011-06-13

    Applicant: George A. Oyler ,  Yien Che Tsai

    Inventor： George A. Oyler ,  Yien Che Tsai

    IPC: C12Q1/37

    CPC classification number: C12Q1/66 ,  C12Q1/37 ,  G01N2333/33 ,  G01N2333/952 ,  G01N2500/10

    Abstract: A cell based assay for detection for protease activity is disclosed. In the assay a cell is engineered to express a protease substrate with at least one label, preferably on its C-terminus. Cleavage of the substrate by the protease that recognizes it results in a C-terminal fragment and a N-terminal fragment, where the fragment having the label is subject to ubiquitin proteasome degradation. The assay measures the disappearance of the label due to degradation of the fragment to which it is attached. A cell free assay is also described for detection of protease activity. In the cell free assay, the protease substrate is expressed in a solution that includes the elements of the ubiquitin proteasome pathway for degradation of the fragment. The assay measures the disappearance of the label attached to the fragment that results from cleavage by the protease.

    Abstract translation: 公开了一种用于蛋白酶活性检测的基于细胞的测定法。 在测定中，将细胞工程化以表达具有至少一个标记的蛋白质底物，优选在其C-末端。 通过识别它的蛋白酶切割底物导致C-末端片段和N-末端片段，其中具有标记的片段经受泛素蛋白酶体降解。 该测定法测量标签由于其附着的片段的降解而消失。 还描述了无细胞测定法用于检测蛋白酶活性。 在无细胞测定中，蛋白酶底物在包含用于降解片段的泛素蛋白酶体途径的元件的溶液中表达。 该测定法测量由蛋白酶切割导致的片段附着的标记物的消失。

    N-END RULE PROTEASE ACTIVITY INDICATION METHODS AND USES THEREOF
    10.
    发明申请
    N-END RULE PROTEASE ACTIVITY INDICATION METHODS AND USES THEREOF 有权
    N末端规则蛋白酶活性指示方法及其用途

    公开(公告)号：US20150010931A1

    公开(公告)日：2015-01-08

    申请号：US13159284

    申请日：2011-06-13

    Applicant: George A. Oyler ,  Yien Che Tsai

    Inventor： George A. Oyler ,  Yien Che Tsai

    IPC: C12Q1/66 ,  C12Q1/37

    CPC classification number: C12Q1/66 ,  C12Q1/37 ,  G01N2333/33 ,  G01N2333/952 ,  G01N2500/10

    Abstract: A cell based assay for detection for protease activity is disclosed. In the assay a cell is engineered to express a protease substrate with at least one label, preferably on its C-terminus. Cleavage of the substrate by the protease that recognizes it results in a C-terminal fragment and a N-terminal fragment, where the fragment having the label is subject to ubiquitin proteasome degradation. The assay measures the disappearance of the label due to degradation of the fragment to which it is attached. A cell free assay is also described for detection of protease activity. In the cell free assay, the protease substrate is expressed in a solution that includes the elements of the ubiquitin proteasome pathway for degradation of the fragment. The assay measures the disappearance of the label attached to the fragment that results from cleavage by the protease.

    Abstract translation: 公开了一种用于蛋白酶活性检测的基于细胞的测定法。 在测定中，将细胞工程化以表达具有至少一个标记的蛋白质底物，优选在其C-末端。 通过识别它的蛋白酶切割底物导致C-末端片段和N-末端片段，其中具有标记的片段经受泛素蛋白酶体降解。 该测定法测量标签由于其附着的片段的降解而消失。 还描述了无细胞测定法用于检测蛋白酶活性。 在无细胞测定中，蛋白酶底物在包含用于降解片段的泛素蛋白酶体途径的元件的溶液中表达。 该测定法测量由蛋白酶切割导致的片段附着的标记物的消失。

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