Proteomic analysis and identification of copper stress-regulated proteins in the marine alga Scytosiphon gracilis (Phaeophyceae)
Proteomic analysis and identification of copper stress-regulated proteins in the marine alga Scytosiphon gracilis (Phaeophyceae)
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DOI：
10.1016/j.aquatox.2009.10.007
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发表时间：
2010-01-31
影响因子：
4.5
通讯作者：
Correa, Juan A.
中科院分区：
文献类型：
--
作者：
Contreras, Loretto;Moenne, Alejandra;Correa, Juan A.
结合肽从头测序和 BLAST 分析的蛋白质组学分析用于鉴定海洋藻类 Scytosiphon gracilis（褐藻纲）中涉及铜耐受性的新型蛋白质。将藻类材料在不含铜（对照）或添加100μg L(-1)的海水中培养4天，通过二维凝胶电泳（2-DE）分离蛋白质提取物。从铜处理获得的蛋白质中，选择25个过表达的蛋白质、5个低表达的蛋白质和5个与对照相比没有变化的蛋白质进行测序。通过毛细管液相色谱和串联质谱 (capLC/MS/MS) 分析从 35 个点获得的胰蛋白酶肽。并通过 BLASTP 确定蛋白质身份。我们鉴定了 19 种过度表达的蛋白质，包括叶绿体过氧化还原酶、胞质磷酸甘露糖变位酶、胞质甘油醛-3-磷酸脱氢酶、3 种 ABC 转运蛋白、伴侣蛋白、蛋白酶体亚基和 tRNA 合酶等。考虑到其他植物模型的可用信息，讨论了这些过度表达的蛋白质可能参与缓冲氧化应激和避免暴露于过量铜的 S. gracilis 中的金属吸收。 (C) 2009 Elsevier B.V. 保留所有权利。
A proteomic analysis combining peptide de novo sequencing and BLAST analysis was used to identify novel proteins involved in copper tolerance in the marine alga Scytosiphon gracilis (Phaeophyceae). Algal material was cultivated in seawater without copper (control) or supplemented with 100 mu g L(-1) for 4 days, and protein extracts were separated by two-dimensional gel electrophoresis (2-DE). From the proteins obtained in the copper treatment, 25 over-expressed, 5 under-expressed and 5 proteins with no changes as compared with the control, were selected for sequencing. Tryptic-peptides obtained from 35 spots were analyzed by capillary liquid chromatography and tandem mass spectroscopy (capLC/MS/MS). and protein identity was determined by BLASTP. We identified 19 over-expressed proteins, including a chloroplast peroxiredoxin, a cytosolic phosphomannomutase, a cytosolic glyceraldehyde-3-phosphate dehydrogenase, 3 ABC transporters, a chaperonine, a subunit of the proteasome and a tRNA synthase, among others. The possible involvement of these over-expressed proteins in buffering oxidative stress and avoiding metal uptake in S. gracilis exposed to copper excess is discussed taking into consideration the information available for other plant models. (C) 2009 Elsevier B.V. All rights reserved.