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    • 6. 发明授权
    • Detection of cellular exposure to ethanol
    • 检测细胞暴露于乙醇
    • US06255057B1
    • 2001-07-03
    • US08686796
    • 1996-07-26
    • Adrienne Sue GordonIvan DiamondDoug Paul Dohrman
    • Adrienne Sue GordonIvan DiamondDoug Paul Dohrman
    • G01N3353
    • G01N33/573G01N33/98G01N2800/307
    • The present invention relates to determinable effects of ethanol exposure on the cellular localization and abundance of specific proteins, referred to herein as ethanol indicative proteins. More specifically, the present invention is based, in part, on the discovery that the catalytic C&agr; subunit of cAMP dependent protein kinase (PKA), which is normally localized in the Golgi apparatus area, appears to translocate to the nucleus upon exposure of a cell to ethanol. The present invention is further based on the observation that the &dgr;-subunit of PKC translocates from the Golgi area to the perinucleus and the nucleus in response to ethanol exposure, while the &egr;-subunit of PKC migrates from the perinucleus into the cytoplasm. The present invention further relates to the discovery that the detectable amount of the regulatory subunit RI of PKA decreases, while the detectable amount of &agr;PKC, &dgr;PKC and &egr;PKC increases upon exposure of a cell to ethanol. These discoveries provide the basis for assays that may be used to detect the exposure of cells to ethanol and assays that may be used for the screening of drugs or the development of treatments to modulate the effects of ethanol consumption. The invention further relates to kits for detecting the exposure of cells to ethanol. Kits of the invention may include antibodies, which preferably are labeled, capable of specifically binding to C&agr; or RI of the cAMP-dependent protein kinase, or, capable of specifically binding to &agr;PKC, &dgr;PKC or &egr;PKC.
    • 本发明涉及乙醇暴露对细胞定位和特异性蛋白质丰度的可确定的影响,本文称为乙醇指示蛋白。 更具体地,本发明部分地基于以下发现:通常位于高尔基体区域中的cAMP依赖性蛋白激酶(PKA)的催化Calpha亚基似乎在细胞暴露时易位于细胞核 到乙醇。 本发明进一步基于以下观察:PKC的δ-亚基从高尔基域转移到响应于乙醇暴露的周核和核,而PKC的ε亚单位从周核迁移到细胞质中。 本发明还涉及PKA的调节亚基RI的可检测量减少的发现,而当细胞暴露于乙醇时,可检测量的αPKC,δPKC和epsiPKC增加。 这些发现提供了可用于检测细胞暴露于乙醇的测定的基础,以及可用于筛选药物或开发用于调节乙醇消耗的影响的处理的测定。 本发明还涉及用于检测细胞暴露于乙醇的试剂盒。 本发明的试剂盒可以包括优选被标记的能够特异性结合cAMP依赖性蛋白激酶的Calpha或RI的抗体,或能够特异性结合αPKC,δPKC或epsiPKC的抗体。