Proteomic analysis of mitochondrial proteins in cardiomyocytes from chronic stressed rat
Proteomic analysis of mitochondrial proteins in cardiomyocytes from chronic stressed rat
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DOI：
10.1002/pmic.200300845
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发表时间：
2004-10-01
期刊：
影响因子：
3.4
通讯作者：
Qian, XH
中科院分区：
文献类型：
--
作者：
Liu, XH;Qian, LJ;Qian, XH
慢性束缚应激可导致心肌细胞损伤，包括严重的超微结构改变和细胞死亡，但其机制和分子基础尚不清楚。线粒体在调节细胞生命中起关键作用。为了探索与应激损伤相关的线粒体蛋白质，应用双向电泳和基质辅助激光解吸/电离质谱（MALDI-TOF MS）。比较慢性束缚应激组和对照组心肌线粒体蛋白质谱，发现11个蛋白质点发生改变，其中7个蛋白质点经MALDI-TOF MS鉴定，其中5个蛋白质点参与慢性束缚应激后线粒体Krebs循环和脂质代谢降低。它们被鉴定为肉毒碱棕榈酰转移酶2、线粒体酰基辅酶A硫酯酶1、异柠檬酸脱氢酶3（NAD+）α、富马酸水合酶1和丙酮酸脱氢酶β。后两种蛋白质，肌酸激酶和抑制素，慢性束缚应激后增加。线粒体能量代谢的生化测试也支持蛋白质组学结果。这些发现为理解慢性应激引起心肌细胞功能障碍或损伤的机制提供了线索。
Chronic restraint stress induces cardiac dysfunction as well as cardiomyocyte injury including severe ultrastructural alteration and cell death, but its mechanism and molecular basis remain unclear. Mitochondria play a key role in regulating cell life. For exploring mitochondrial proteins which correlate with stress-induced injury, two-dimensional electrophoresis and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) were applied. After comparing the protein profiles of myocardial mitochondria between a chronic restraint stress group and a control group, 11 protein spots were found altered, seven of which were identified by MALDI-TOF MS. Among the seven proteins, five proteins involved in the Krebs cycle and lipid metabolism in mitochondria decreased after chronic restraint stress. They were identified as carnitine palmitoyltransferase 2, mitochondrial acyl-CoA thioesterase 1, isocitrate dehydrogenase 3 (NAD+) alpha, fumarate hydratase 1 and pyruvate dehydrogenase beta. The last two proteins, creatine kinase and prohibitin, increased after chronic restraint stress. Biochemical tests for energy metabolism in mitochondria also supported the proteomic results. These findings provide clues for understanding the mechanism of dysfunction or injury in cardiomyocytes induced by chronic stress.