Effects of oxidative stress on the cardiac myocyte proteome: modifications to peroxiredoxins and small heat shock proteins
Effects of oxidative stress on the cardiac myocyte proteome: modifications to peroxiredoxins and small heat shock proteins
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DOI：
10.1016/j.yjmcc.2005.10.004
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发表时间：
2006-01-01
影响因子：
5
通讯作者：
Sugden, PH
中科院分区：
文献类型：
--
作者：
Cullingford, TE;Wait, R;Sugden, PH
内源性氧化应激可能是体内心肌细胞死亡的原因。我们研究了早期（0-2小时）暴露于过氧化氢（0.1 mM）的新生大鼠分离的心肌细胞的蛋白质组的变化，集中在蛋白质的表观分子量为20和30 kDa之间。蛋白质通过双向凝胶电泳（2DGE）分离，通过银染定位和质谱鉴定。还研究了[S-35]蛋氨酸或（32）Pi的掺入。对于选定的蛋白质，通过逆转录酶-聚合酶链反应检查转录丰度。在该区域的38个蛋白质点中，鉴定了23个。两个家庭表现出变化的2DGE迁移或丰富的H2 O2处理：过氧化物酶和两个小的热休克蛋白（Hsp）家族成员：热休克27 kDa蛋白1（Hsp 25）和α B-晶状体蛋白。过氧化物酶转移到较低的pI值，这可能是由于活性位点半胱氨酸残基的“过度氧化”。热休克蛋白25也转移到较低的pI值，但这是由于磷酸化。α B-晶状体蛋白迁移没有变化，但其丰度降低。转录本编码过氧化物酶2和5显着增加。此外，还鉴定了另外10种蛋白质。对于两种（谷胱甘肽S-转移酶pi，免疫控制的肿瘤蛋白），我们无法找到任何先前的参考文献表明它们在心肌细胞中的发生。我们的结论是，暴露于心肌细胞的氧化应激导致两个蛋白质家族参与细胞保护的翻译后修饰。这些变化可能在诊断上是有用的。在短期内，氧化应激引起的全球蛋白质丰度的变化很少检测到银染色评估。(c)2005爱思唯尔有限公司保留所有权利。
Endogenous oxidative stress is a likely cause of cardiac myocyte death in vivo. We examined the early (0-2 h) changes in the proteome of isolated cardiac myocytes from neonatal rats exposed to H2O2 (0.1 mM), focussing on proteins with apparent molecular masses of between 20 and 30 kDa. Proteins were separated by two-dimensional gel electrophoresis (2DGE), located by silver-staining and identified by mass spectrometry. Incorporation of [S-35] methionine or (32)Pi was also studied. For selected proteins, transcript abundance was examined by reverse transcriptase-polymerase chain reaction. Of the 38 protein spots in the region, 23 were identified. Two families showed changes in 2DGE migration or abundance with H2O2 treatment: the peroxiredoxins and two small heat shock protein (Hsp) family members: heat shock 27 kDa protein 1 (Hsp25) and alpha B-crystallin. Peroxiredoxins shifted to lower pI Values and this was probably attributable to 'over-oxidation' of active site Cys-residues. Hsp25 also shifted to lower pI values but this was attributable to phosphorylation. alpha B-crystallin migration was unchanged but its abundance decreased. Transcripts encoding peroxiredoxins 2 and 5 increased significantly. In addition, 10 further proteins were identified. For two (glutathione S-transferase pi, translationally-control led tumour protein), we could not find any previous references indicating their occurrence in cardiac myocytes. We conclude that exposure of cardiac myocytes to oxidative stress causes post-translational modification in two protein families involved in cytoprotection. These changes may be potentially useful diagnostically. In the short term, oxidative stress causes few detectable changes in global protein abundance as assessed by silver-staining. (c) 2005 Elsevier Ltd. All rights reserved.