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Caspase-3体外对PIAS1蛋白酶切作用的初步验证
赵瑞1,唐春花1,赵文元1,刘建民1*,洪波1,许奕1,黄清海1,霍克克2
0
(1. 第二军医大学长海医院临床神经医学中心,神经外科,上海 200433
2. 复旦大学生命科学学院遗传学研究所,上海 200433
*通信作者)
摘要:
目的 观察凋亡相关蛋白Caspase-3体外对PIAS1蛋白的酶切作用,为后续研究奠定基础。方法 以HEK293T细胞表达PIAS1蛋白,采用免疫共沉淀(Co-IP)实验观察其与 Caspase-3间的结合作用;采用体外蛋白酶切实验检测Caspase-3对PIAS1的酶切作用。结果 Co-IP实验发现体外Caspase-3可与PIAS1蛋白结合;蛋白酶切实验发现PIAS1的酶切片段。结论 Caspase-3与PIAS1在体外具有相互作用,PIAS1体外可被Caspase-3酶切,为后续研究奠定了基础。
关键词:  Caspase-3  PIAS1  酶切;体外
DOI:10.3724/SP.J.1008.2011.01144
投稿时间:2011-05-29修订日期:2011-10-08
基金项目:国家自然科学基金项目
Cleavage effect of Caspase-3 on PIAS1 protein in vitro
ZHAO Rui1,TANG Chun-hua1,ZHAO Wen-yuan1,LIU Jian-min1*,HONG Bo1,XU Yi1,HUANG Qing-hai1,HUO Ke-ke2
(1. Department of Neurosurgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
2. Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China
*Corresponding author.)
Abstract:
Objective To observe the cleavage effect of Caspase-3 on PIAS1 protein in vitro. Methods PIAS1 protein was expressed in HEK293T cells. Co-immunoprecipitation (Co-IP) was used to observe the binding between PIAS1 protein and Caspase-3. The cleavage effect of Caspase-3 on PIAS-1 protein was detected by in vitro proteolytic cleavage test. Results Co-IP result showed that Caspase-3 bond to PIAS1 in vitro. Proteolytic cleavage test showed that PIAS1 was cleaved by active Caspase-3. Conclusion It has been shown that Caspase-3 can interact with PIAS1 protein in vitro, and PIAS1 can be cleaved by Caspase-3 in vitro.
Key words:  Caspase-3  PIAS1  cleavage