低氧预适应小鼠脑皮层内与nPKCε相互作用蛋白的鉴定  

Identification of nPKCε-interacted proteins in the cortex of hypoxic preconditioned mice

作　　者：封素娟[1] 刘旭[1] 张彩艳[1] 卜祥宁[1] 张楠[1] 孙媛 郭菲 李俊发[1] 

机构地区：[1]首都医科大学神经生物学系北京神经科学研究所,北京100069 [2]北京市科协

出　　处：《基础医学与临床》2010年第3期225-231,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(30670782;30871219);国家"973"计划(2006CB5041);北京市自然科学基金A类项目(5072008);北京市教育委员会科技计划重点项目(KZ200810025012);北京市属高等学校人才强教计划项目(PHR200906116);北京市新世纪百千万人才工程培养经费(08-016)

摘　　要：目的鉴定脑低氧预适(HPC)小鼠脑皮层组织内与新奇型蛋白激酶Cε(nPKCε)相互作用的蛋白。方法利用免疫沉淀(IP)和双向电泳(2-DE)技术分离小鼠脑皮层中与nPKCε相互作用的蛋白;以ImageMaster2D Platinum软件对双向电泳图谱进行差异表达分析;目标蛋白用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)及蛋白印迹(Western blot)进行鉴定分析。结果与对照组比较,HPC小鼠脑皮层内与nPKCε相互作用的胞质成分中有34个蛋白点上调,20个蛋白点下调,膜相关成分中有27个蛋白点上调,28个蛋白点下调;HPC后脑皮层胞质及膜相关成分中与nPKCε相互作用的热休克蛋白(HSP)70和14-3-3γ的蛋白表达量均升高,而HSP60仅在膜相关成分中升高(P<0.05,n=3)。结论nPKCε可能通过调节HSP60、HSP70和14-3-3γ等多种与其相互作用的蛋白参与脑HPC的形成。Objective Identify novel protein kinase Cε（nPKCε）-interacted proteins in the cortex of hypoxic preconditioned mice.Methods Immunoprecipitation （IP） and two-dimensional electrophoresis（2-DE） combining with ImageMaster 2D Platinum software were applied to analyze the differential expressions of nPKCε-interacted proteins;the target protein spots were identified by matrix-associated laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and Western blot.Results Compared with control group,there were 34 upregulated protein spots and 20 downregulated protein spots in cytosolic fraction,while 27 upregulated prtein spots and 28 downregulated protein spots were determined in particulate fraction of cerebral cortex of HPC mice.The levels of nPKCε-interacted HSP70 and 14-3-3γ protein expressions increased significantly in both cytosolic and particulatefractions;but the protein level of nPKCε-interacted HSP60 increased only in particulate fraction of cerebral cortex of HPC mice.Conclusion nPKCε might be involved in the development of cerebral HPC via the regulation of its interacted proteins such as HSP60,HSP70 and 14-3-3γ.

关 键 词：低氧预适应 新奇型蛋白激酶Cε 蛋白质组学 蛋白表达量 

分 类 号：R741[医药卫生—神经病学与精神病学]