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    Isozyme of autoclavable superoxide dismutase (SOD), a process for the identification and extraction of the SOD in cosmetic, food and pharmaceutical compositions
    123.
    发明授权
    Isozyme of autoclavable superoxide dismutase (SOD), a process for the identification and extraction of the SOD in cosmetic, food and pharmaceutical compositions 有权
    高压灭菌超氧化物歧化酶（SOD）的同功酶，用于鉴定和提取化妆品，食品和药物组合物中的SOD的方法

    公开(公告)号：US06485950B1

    公开(公告)日：2002-11-26

    申请号：US09617118

    申请日：2000-07-14

    Applicant: Sanjay Kumar ,  Rashmita Sahoo ,  Paramvir Singh Ahuja

    Inventor： Sanjay Kumar ,  Rashmita Sahoo ,  Paramvir Singh Ahuja

    IPC: C12N902

    CPC classification number: A23G4/123 ,  A61K8/66 ,  A61K38/00 ,  A61Q19/00 ,  C12N9/0089 ,  C12Y115/01001 ,  Y10S977/915 ,  Y10S977/926

    Abstract: The invention relates to a novel purified isozyme of an autoclavable superoxide dismutase extracted from the plant Potentilla atrosanguinea Lodd. Var. orgyrophylla, said isozyme having the following characteristics, O2− scavenging activity remains same before and after autoclaving; scavenges O2− from sub-zero temperature of −20° C. to high temperature of +80° C.; O2− scavenging activity at 25° C. for 30 days without adding any stabilizing agent such as polyols or sugars; O2− scavenging activity in the presence of saline (0.9% sodium chloride) to 61.8% of the control (without 0.9% sodium chloride), stable at 4° C. for at least 8 months; contamination free and infection free from any living micro- and/or macro-organism after autoclaving; possesses temperature optima at 0° C.; possesses a molecular weight of 33 kD under non-denaturating conditions; possesses a molecular weight of 36 kD under denaturating conditions; has clear peaks in UV range at 268 and 275 nm; has an enzyme turnover number of 19.53×104% per nmol per min at 0° C.; and requires Cu/Zn as a co-factor, method for the preparation of the purified isozyme of autoclavable superoxide dismutase and formulations containing the said autoclavable superoxide dismutase.

    Abstract translation: 本发明涉及从植物Potentilla atrosanguinea Lodd提取的可高压灭菌的超氧化物歧化酶的新型纯化同功酶。 可变 所述同功酶具有以下特征，在高压灭菌之前和之后，O2-清除活性保持相同; 清除O2-从零度温度-20°C到+ 80°C的高温。 氧气清除活性在25℃下30天，而不添加任何稳定剂如多元醇或糖; 在盐水（0.9％氯化钠）存在下，将清除氧气的活性提高到对照组（不含0.9％氯化钠）的61.8％，4℃稳定至少8个月; 高压灭菌后无任何生物微生物和/或大生物感染的免污染和感染; 在0°C时具有最佳温度。 在非变性条件下具有33 kD的分子量; 在变性条件下具有36kD的分子量; 在268和275nm的UV范围内具有清晰的峰; 在0℃下每分钟酶转化数为19.53×104％/ nmol。 并且需要Cu / Zn作为辅助因子，制备可高压灭菌的超氧化物歧化酶的纯化的同功酶的方法和含有所述高压灭菌的超氧化物歧化酶的制剂。

    Methods of the identification of pharmaceutically active compounds
    124.
    发明授权
    Methods of the identification of pharmaceutically active compounds 有权
    鉴定药物活性化合物的方法

    公开(公告)号：US06218136B1

    公开(公告)日：2001-04-17

    申请号：US09142551

    申请日：1998-09-10

    Applicant: Sanjay Kumar ,  George Pietro Livi ,  Megan McHale McLaughlin ,  Peter Ronald Young

    Inventor： Sanjay Kumar ,  George Pietro Livi ,  Megan McHale McLaughlin ,  Peter Ronald Young

    IPC: C12Q148

    CPC classification number: C12N9/1205 ,  A61K38/00 ,  A61K49/0004 ,  C07K2319/00 ,  C12Q1/485 ,  G01N33/573 ,  G01N33/6863

    Abstract: CSBP/p38 is a MAP kinase that is activated in response to stress, endotoxin, interleukin 1 and tumor necrosis factor. Using a catalytically inactive mutant (D168A) of human CSBP2 as the bait in a yeast two-hybrid screen, a kinase has been cloned which shares ˜70% amino acid identity to MAPKAP kinase-2, and thus was designated MAPKAP kinase-3. The binding of CSBP to MAPKAP kinase 3 was confirmed in vitro by the precipitation of epitope-tagged CSBP1, CSBP2 and CSBP2(D168A) and endogenous CSBP from mammalian cells by a bacterially-expressed GST-MAPKAP kinase-3 fusion protein and in vivo by co-precipitation of the epitope-tagged proteins co-expressed in HeLa cells. MAPKAP kinase-3 was phosphorylated by both CSBP1 and CSBP2, and was then able to phosphorylate HSP27 in vitro. Treatment of HeLa cells with sorbitol or TNF resulted in activation of CSBP and MAPKAP kinase-3 and activation of MAPKAP kinase-3 could be blocked by preincubation of cells with 4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-4-pyridyl)-1H-imidazole, a specific inhibitor of CSBP kinase activity. These data suggest that MAPKAP kinase-3 is activated by stress and cytokines and is a novel substrate of CSBP both in vitro and in vivo. The use of MAPKAP kinase-3 in screens for the identification of pharmaceutically active compounds is disclosed.

    Abstract translation: CSBP / p38是一种MAP应激反应的激酶，内毒素，白细胞介素1和肿瘤坏死因子。 在酵母双杂交筛选中使用人CSBP2的催化无活性突变体（D168A）作为诱饵，已经克隆了与MAPKAP激酶-2共享〜70％氨基酸同一性的激酶，因此被命名为MAPKAP激酶-3。 通过细菌表达的GST-MAPKAP激酶-3融合蛋白从哺乳动物细胞中沉淀表位标记的CSBP1，CSBP2和CSBP2（D168A）和内源性CSBP，体外证实CSBP与MAPKAP激酶3的结合 在HeLa细胞中共表达的表位标记蛋白的共沉淀。 MAPKAP激酶-3被CSBP1和CSBP2磷酸化，然后能够在体外磷酸化HSP27。 用山梨糖醇或TNF处理HeLa细胞导致CSBP和MAPKAP激酶-3的活化，MAPKAP激酶-3的激活可以通过用4-（4-氟苯基）-2-（4-甲基亚磺酰基苯基）-5 -4-吡啶基）-1H-咪唑，CSBP激酶活性的特异性抑制剂。 这些数据表明MAPKAP激酶-3被应激和细胞因子激活，并且是体外和体内CSBP的新型底物。 公开了在筛选中使用MAPKAP激酶-3鉴定药物活性化合物。

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