| 张枫,孟祥兵,宋宜,梅柱中,董燕,刘斌,孙志贤.cAMP-PKA参与辐射诱导p27Kip1蛋白的表达调控[J].中华放射医学与防护杂志,2004,24(5):393-395 |
| cAMP-PKA参与辐射诱导p27Kip1蛋白的表达调控 |
| Involvement of cAMP-PKA in regulation of p27Kip1 expression induced by ionizing radiation |
| 投稿时间：2004-01-05 |
| DOI： |
| 中文关键词: 电离辐射 p27Kip1 PKA 蛋白磷酸化 |
| 英文关键词:Ionizing radiation p27Kip1 PKA Protein phosphorylation |
| 基金项目:国家自然科学基金资助项目(30070239;30170291) |
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| 中文摘要: |
| 目的 研究电离辐射反应中p27Kip1表达调控机理。方法 将p27Kip1蛋白编码全长基因克隆导入原核表达载体pGEX4T-2,经异丙基硫代-β-D半乳糖苷(IPTG)诱导表达后再经GST-Sephrose4B柱亲和层析、纯化,获取的GST-p27Kip1融合蛋白作为磷酸化底物,并进行体外磷酸化检测。结果 PKA具有磷酸化p27Kip1蛋白的功能,该反应能被PKA的特异性抑制剂H89所抑制;照射后细胞内源性的PKA对p27Kip1的磷酸化程度减弱。结论 cAMPPKA在电离辐射诱导细胞周期检查点蛋白p27Kip1表达降低的调控机理中起到主要作用。 |
| 英文摘要: |
| Objective To study the mechanism of p27Kip1 down-regulation induced by ionizing radiation(IR). Methods The cDNA of p27Kip1 was cloned into prokaryotic expression vector pGEX4T-2.The recombinant protein GST-p27Kip1 was induced by isopropyl-β-D-thiroglactoside (IPTG) and purified by GST-Sepharose 4B affinity chromatography. Results GST-p27Kip1 was phosphorylated by PKAc,and this in vitro phosphorylation could be inhibited by PKA specific inhibitor H89.Phosphorylation level of GST-p27Kip1 decreased after IR,when treated with immunoprecipited endogenous PKA. Conclusion The results suggest that cAMP-PKA plays a critical role in down-regulation of p27Kip1 during IR. |
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