Myocardial subproteomic analysis of a constitutively active Rac1-expressing transgenic mouse with lethal myocardial hypertrophy
Myocardial subproteomic analysis of a constitutively active Rac1-expressing transgenic mouse with lethal myocardial hypertrophy
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DOI：
10.1152/ajpheart.01041.2004
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发表时间：
2005-12-01
影响因子：
4.8
通讯作者：
Van Eyk, JE
中科院分区：
文献类型：
--
作者：
Buscemi, N;Murray, C;Van Eyk, JE
一个具有致死性心肌肥大的组成型活性Rac 1表达转基因小鼠的心肌亚蛋白质组学分析。Am J Physiol Heart Circ Physiol 289：H2325-H2333，2005; doi：10.1152/ajpheart.01041.2004。- 一个二维凝胶电泳（2-DE）为基础的蛋白质组学方法被用来研究一个小的单体GTdR，Rac 1，组成型表达只在心肌中的acerbated扩张型心肌病的转基因小鼠模型。亚分级程序允许集中分析的细胞质和肌丝蛋白富集提取物的心室组织从Rac 1转基因和年龄匹配的非转基因（NTG）小鼠。这些小鼠中的大多数表现出严重的肥大（Rac 1小鼠的心脏与体重比> 2倍），并在第14天和第17天之间死于明显的心力衰竭。来自Rac 1（n = 4）和NTG（n = 4）组的比较性2-DE分析（pH 3-10，12% SDS-PAGE）揭示了平均蛋白质斑点强度的差异。来自细胞质蛋白富集提取物的12种蛋白质符合我们的稳健性和斑点分辨率标准，并进行了鉴定。这些蛋白质代表了细胞功能的广泛分布，只有一些先前涉及心肌肥大。肌丝亚蛋白质组在翻译后修饰中没有变化，但通过一维Western印迹的进一步分析显示Rac 1小鼠心室中肌丝蛋白的数量增加。此外，在第7、9和11天分析了在细胞质蛋白富集亚蛋白质组中改变的三种具有不同功能的蛋白质，微管蛋白β链、锰超氧化物歧化酶和苹果酸脱氢酶，以评估它们在扩张型心肌病表型发展中的作用。所有三种蛋白质的量在第9天达到峰值，表明在心脏肥大性衰竭的早期反应。
Myocardial subproteomic analysis of a constitutively active Rac1-expressing transgenic mouse with lethal myocardial hypertrophy. Am J Physiol Heart Circ Physiol 289: H2325-H2333, 2005; doi:10.1152/ajpheart.01041.2004. - A two-dimensional gel electrophoresis (2-DE)- based proteomic approach was used to study a transgenic mouse model of acerbated dilated cardiomyopathy in which the small monomeric GTPase, Rac1, was constitutively expressed exclusively in the myocardium. A subfractionation procedure allowed for the focused analysis of both cytoplasmic and myofilament protein-enriched extracts of ventricular tissue from Rac1 transgenic and age-matched nontransgenic (NTG) mice. The majority of these mice displayed severe hypertrophy (heart-to-body weight ratios > 2-fold greater in the Rac1 mice) and died from overt heart failure between days 14 and 17. Comparative 2-DE analysis (pH 3-10, 12% SDS-PAGE) derived from Rac1 (n = 4) and NTG (n = 4) groups revealed differences in mean protein spot intensities. Twelve proteins from the cytoplasmic protein-enriched extract met our criteria for robustness and spot resolution and were identified. These proteins represent a broad distribution of cellular functions with only some previously implicated in myocardial hypertrophy. The myofilament subproteome displayed no change in posttranslational modification, but further analysis by one-dimensional Western blot showed increased quantities of myofilament proteins in the Rac1 mouse ventricles. Additionally, three proteins with different functionality that were altered in the cytoplasmic protein-enriched subproteome, tubulin beta-chain, manganese superoxide dismutase, and malate dehydrogenase, were analyzed at days 7, 9, and 11 to assess their role in the development of the dilated cardiomyopathic phenotype. The quantity of all three proteins peaked at day 9, suggesting an early response in cardiac hypertrophic failure.