Protein carbonyl formation in the diaphragm
Protein carbonyl formation in the diaphragm
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DOI：
10.1165/rcmb.2004-0021oc
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发表时间：
2005-01-01
影响因子：
6.4
通讯作者：
Hussain, SNA
中科院分区：
文献类型：
--
作者：
Barreiro, E;Gea, J;Hussain, SNA
虽然蛋白质羰基形成是骨骼肌氧化应激的一个指标，但在这些肌肉中发生氧化的确切蛋白质仍不清楚。我们使用二维电泳、免疫印迹和质谱法鉴定脓毒症动物膈膜中的羰基化蛋白。给大鼠注射生理盐水（对照）或大肠杆菌脂多糖（LPS），并在不同时间间隔后处死。肌膜蛋白羰基化在注射LPS后12 h达到峰值，且羰基化在肌纤维内和供应肌纤维的血管内均有发生。醛缩酶A、甘油醛3-磷酸脱氢酶、烯醇化酶3 β、线粒体和胞质肌酸激酶、α -肌动蛋白、碳酸酐酶III和泛醇细胞色素c还原酶在脓毒症大鼠膈膜中均羰基化。此外，我们发现羰基化强度与肌酸激酶和醛缩酶活性之间存在显著的负相关。我们得出结论，在败血症期间，糖酵解、ATP生产、CO2水合作用和收缩蛋白是膈膜内氧自由基的目标。
Although protein carbonyl formation is an index of oxidative stress in skeletal muscles, the exact proteins, which undergo oxidation in these muscles, remain unknown. We used 2D electrophoresis, immunoblotting, and mass spectrometry to identify carbonylated proteins in the diaphragm in septic animals. Rats were injected with saline (control) or Escherichia coli lipopolysaccharides (LPS) and killed after various intervals. Diaphragm protein carbonylation increased significantly and peaked 12 h after LPS injection, and it was localized both inside muscle fibers and in blood vessels supplying muscle fibers. Aldolase A, glyceraldehyde 3-phosphate dehydrogenase, enolase 3beta, mitochondrial and cytosolic creatine kinases, alpha-actin, carbonic anyhdrase III, and ubiquinol-cytochrome c reductase were all carbonylated in septic rat diaphragms. In addition, we found significant negative correlations between the intensity of carbonylation and creatine kinase and aldolase activities. We conclude that glycolysis, ATP production, CO2 hydration, and contractile proteins are targeted by oxygen radicals inside the diaphragm during sepsis.