CAJ | 학술논문

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Ce(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nPKCe相互作用的蛋白;以ImageMaster 2D 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nPKCe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 nPKCe可能通过调节HSP60、HSPT0和14-3-3γ等多种与其相互作用的蛋白参与脑HPC的形成.
목적 감정뇌저양예괄(HPC)소서뇌피층조직내여신기형단백격매Ce(nPKCε)상호작용적단백.방법 이용면역침정(IP)화쌍향전영(2-DE)기술분리소서뇌피층중여nPKCe상호작용적단백;이ImageMaster 2D Platinum 연건대쌍향전영도보진행차이표체분석;목표단백용기질보조격광해석전리비행시간질보(MALDI-TOF-MS)급단백인적(Western blot)진행감정분석.결과 여대조조비교,HPC소서뇌피층내여nPKCε상호작용적포질성분중유34개단백점상조,20개단백점하조,막상관성분중유27개단백점상조,28개단백점하조;HPC후뇌피층포질급막상관성분중여nPKCe상호작용적열휴극단백(HSP)70화14-3-3γ적단백표체량균승고,이HSP60부재막상관성분중승고(P＜0.05,n=3).결론 nPKCe가능통과조절HSP60、HSPT0화14-3-3γ등다충여기상호작용적단백삼여뇌HPC적형성.
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 nPKCe-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 HSP 70 and 14-3-3γ/protein expressions increased significantly in both cytosolic and particulate fractions;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.