Analysis of oligomeric protein complexes in the chloroplast sub-proteome of nucleic acid-binding proteins from mustard reveals potential redox regulators of plastid gene expression
Analysis of oligomeric protein complexes in the chloroplast sub-proteome of nucleic acid-binding proteins from mustard reveals potential redox regulators of plastid gene expression
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DOI：
10.1002/pmic.200900678
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发表时间：
2010-06-01
期刊：
影响因子：
3.4
通讯作者：
Pfannschmidt, Thomas
中科院分区：
文献类型：
--
作者：
Schroeter, Yvonne;Steiner, Sebastian;Pfannschmidt, Thomas
植物光合光质驯化涉及氧化还原控制的质体基因表达变化。为了研究可能参与这一调控的蛋白质，我们从十字花菜（Sinapis alba）中分离出低丰度的叶绿体核酸结合蛋白，并研究光合氧化还原信号是否影响其组成和/或寡聚结构。我们从遭受光质量变化的植物中纯化叶绿体，并将细胞器裂解物应用于肝素- sepharose层析，然后使用二维蓝色原生PAGE。我们研究了低聚蛋白复合物的积累和结构，并应用质谱联用技术对其进行鉴定。我们发现了10个高阶寡聚蛋白复合物和11个较小的蛋白复合物或点，包括质体编码RNA聚合酶（PEP）、质体转录活性染色体蛋白、RNA结合蛋白、核糖体亚基和伴侣蛋白。翻译延伸因子被发现是唯一的蛋白质，在其数量上显示出主要差异，以响应生长光。此外，我们还发现了一种新的硫氧还蛋白作为PEP的亚基，一种2- cys -过氧化物还蛋白复合物和一种（可溶性）铁氧还蛋白：nadp氧化还原酶，它们代表了质体基因表达的潜在氧化还原调节因子。来自拟南芥的硫氧还蛋白的T-DNA敲除线显示出淡黄色表型，表明这种新的PEP亚基对于质体的正常发育至关重要。
Photosynthetic light quality acclimation in plants involves redox-controlled changes in plastid gene expression. To study proteins potentially involved in this regulation, we isolated low-abundant chloroplast nucleic acid-binding proteins from the crucifere mustard (Sinapis alba) and investigated if photosynthetic redox signals affect their composition and/or oligomeric structure. We purified chloroplasts from plants subjected to light quality shifts and applied organelle lysates to heparin-Sepharose chromatography followed by 2-D blue native PAGE. We studied accumulation and structure of oligomeric protein complexes and applied MS/MS to identify them. We found ten oligomeric protein complexes of higher order and eleven smaller protein complexes or spots including plastid-encoded RNA polymerase (PEP), plastid transcriptionally active chromosome proteins, RNA-binding proteins, ribosomal subunits and chaperones. A translation elongation factor was found to be the only protein displaying major differences in its amounts in response to the growth lights. Furthermore, we found a novel thioredoxin as a subunit of the PEP, a 2-Cys-peroxiredoxin complex and a (soluble) ferredoxin:NADP-oxido-reductase, which represent potential redox regulator of plastid gene expression. A T-DNA knock-out line of the thioredoxin from Arabidopsis exhibits a yellowish-pale phenotype, demonstrating that this novel PEP subunit is essential for proper plastid development.