高原鼠兔和Wistar大鼠骨骼肌线粒体的比较蛋白质组学初步研究

目的 采用比较蛋白质组学方法研究高原鼠兔和Wistar大鼠线粒体蛋白的差异.方法 分别在海拔4 500 m高原(高原鼠兔)和模拟4 500 m低压舱内麻醉动物后取双侧腓肠肌,分离纯化线粒体蛋白进行二维电泳和凝胶图像分析,对差异蛋白质用MALDI-TOF-MS质谱仪进行肽指纹图谱测定和ESI-Q-TOF串联质谱鉴定.结果 高原鼠兔和Wistar大鼠线粒体蛋白二维凝胶图谱有明显差异;鉴定了泛醌-细胞色素C还原酶核心蛋白Ⅰ、硫氧还原蛋白依赖性过氧化物还原酶线粒体前体蛋白、线粒体胸苷激酶2、肌酸激酶线粒体异构体4、推测蛋白XP_499518、未知功能蛋白、肌酸激酶、F1F0-ATP酶d亚基、镁依赖性蛋白磷酸酶1D、推测蛋白LOC498909等差异线粒体蛋白.结论 高原鼠兔和Wistar大鼠有差异线粒体蛋白,提示高原鼠兔线粒体能量代谢、抗氧化能力、线粒体DNA稳定性等方面有其特点.

Comparative proteomic analysis of plateau Pika and Wistar rat gastrocnemius mitochondria

Objective To explore high-altitude-adaptation-related mitochondrial proteins by analyzing the differences of gastrocnemius mitochondrial protein profiles between plateau Pika and Wistar rat. Methods The plateau Pikas were captured in Kunlun Mountain (4 500 m) while Wistar rats were bred at low altitude (Chongqing, 300 m) and exposed to hypobaric hypoxia (4 500 m) for 30 d. After animals of both groups were anesthetized, bilateral gastrocnemius were collected and mitochondria were extracted and purified. Two dimensional electrophoresis and PDQuest were applied and the peptide mass fingerprinting of differential proteins was analyzed with matrix assisted laser desorption/ionization mass spectrometry analysis (MALDI-TOF MS) and Q-TOF(quatropde-time of flight)MS/MS. Results Great difference in gastrocnemius mitochondrial profiles of plateau Pika and Wistar rat was found. Among the differential protein spots, 11 were identified. Six plateau Pika specific protein spots were identified as ubiquinol-cytochrome c reductase core protein I, creatine kinase, mitochondrial isoform 4, glyceraldehyde-3-phosphate dehydrogenase, creatine kinase, hypothetical protein XP_499518 and a novel protein. Five Wistar rat specific protein spots were identified as subunit d of mitochondrial H-ATP synthase (2 spots), LOC498909 protein and creatine kinase. Conclusion Our results suggest the difference in mitochondrial energy metabolism, antioxidative activity, mitochondrial DNA stability exist between plateau Pika and Wistar rat.