Proteomic analysis of cisplatin-induced cochlear damage: Methods and early changes in protein expression
Proteomic analysis of cisplatin-induced cochlear damage: Methods and early changes in protein expression
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DOI：
10.1016/j.heares.2006.12.017
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发表时间：
2007-04-01
期刊：
影响因子：
2.8
通讯作者：
Salvi, Richard J.
中科院分区：
文献类型：
--
作者：
Coling, Donald E.;Ding, Dalian;Salvi, Richard J.
为了确定与顺铂耳毒性相关的蛋白质表达的早期变化，我们使用二维差异凝胶电泳（2D-DIGE）和基质辅助激光解吸飞行时间（MALDI-TOF）质谱分析蛋白质从P3大鼠耳蜗培养3小时或没有1 mM顺铂。对来自6个凝胶的荧光图像的重复分析揭示了顺铂诱导的22种耳蜗蛋白表达的显著变化（p < 0.01）（大于1.5倍）。这些包括五种蛋白质表达的增加，其中四种被鉴定为核结合蛋白1，一种核钙信号和稳态蛋白。（2.1倍），异质核核糖核蛋白C，RNA加工蛋白（1.8倍），一种55 kDa的蛋白质，是内皮分化相关因子1或α-6微管蛋白（1.7倍）和钙网蛋白，内质网的钙结合伴侣（ER，1.6倍）。17种蛋白质的表达显著（p < 0.01）降低了1.5倍以上。这些包括核糖核酸酶/血管生成素抑制剂1（1.6倍），RAS样，家族12（预测），ras相关（RaIGDS/AF-6）结构域家族5（4.5倍），同源的RAS家族的GT3信号蛋白（2.4倍），和蛋白酪氨酸磷酸酶结构域包含1（预测，6.1倍）。我们鉴定了7种耳蜗蛋白，它们在顺铂诱导的表达变化中具有较小（1.2-1.5倍）或不太显著（p < 0.05）的变化。值得注意的是，热休克70 kDa蛋白5（Hspa 5，Grp 78和BiP），一种参与应激反应的ER伴侣蛋白，减少了1.7倍。我们观察到的变化与另一种ER应激诱导蛋白，葡萄糖调节蛋白Grp 58的亚型水平的磷酸化一致。顺铂诱导的蛋白质表达的变化进行了讨论，相对于已知的或假设的功能，所确定的蛋白质。(C)2007 Elsevier B. V.保留所有权利。
To identify early changes in protein expression associated with cisplatin ototoxicity, we used two dimensional-difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption-time-of-flight (MALDI-TOF) mass spectrometry to analyze proteins from P3 rat cochleae that were cultured for 3 h with or without 1 mM cisplatin. Replicate analysis of fluorescent images from six gels revealed significant (p < 0.01) cisplatin-induced changes (greater than 1.5-fold) in expression of 22 cochlear proteins. These include increases in the expression of five proteins, four of which were identified as nucleobindin 1, a nuclear calcium signaling and, homeostasis protein (2.1-fold), heterogeneous nuclear ribonucleoprotein C, an RNA processing protein (1.8-fold), a 55 kDa protein that is either endothelial differentiation-related factor 1 or alpha-6 tubulin (1.7-fold), and calreticulin, a calcium binding chaperone of the endoplasmic reticulum (ER, 1.6-fold). The expression of 17 proteins was significantly (p < 0.01) decreased by greater than 1.5-fold. These include ribonuclease/ angiogenin inhibitor 1 (1.6-fold), RAS-like, family 12 (predicted), ras association (RaIGDS/AF-6) domain family 5 (4.5-fold), homologous the RAS family of GTPase signaling proteins (2.4-fold), and Protein tyrosine phosphatase domain containing 1 (predicted, 6.1-fold). We identified seven cochlear proteins with either smaller (1.2-1.5-fold) or less significant (p < 0.05) cisplatin-induced changes in expression. Notably, heat shock 70 kDa protein 5 (Hspa5, Grp78, and BiP), an ER chaperone protein involved in stress response, decreased 1.7-fold. We observed changes consistent with phosphorylation in the level of isoforms of another ER stress-induced protein, glucose-regulated protein Grp58. Changes in cisplatin-induced protein expression are discussed with respect to known or hypothesized functions of the identified proteins. (C) 2007 Elsevier B.V. All rights reserved.