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首页> 外文期刊>Journal of Experimental Botany >Grape berry plasma membrane proteome analysis and its differential expression during ripening
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Grape berry plasma membrane proteome analysis and its differential expression during ripening

机译：葡萄浆果质膜蛋白质组学分析及其在成熟过程中的差异表达

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High purity berry plasma membranes (PMs) of Vitis vinifera L. cv. Cabernet Sauvignon were isolated by two-phase partitioning of microsome fractions at different stages of berry ripening. PM proteins resolvable by the detergent cocktail of CHAPS and ASB-14 were separated by two-dimensional electrophoresis. A total of 119 protein spots from pre-véraison berry PMs on 2-D gels detected with silver staining were subjected to MALDI-TOF mass spectrometry analysis. Sixty-two spots were identified as putative PM proteins, with 1–6 predicted transmembrane helices, including true PM proteins such as ATP synthase, ABC transporters, and GTP-binding proteins reported in plants. They were then grouped into eight functional categories, mainly involved in transport, metabolism, signal transduction, and protein synthesis. Another 11 spots were identified as proteins of unknown function. The véraison and post-véraison samples stained 98 and 86 spots on the gels, respectively. During the berry ripening process, total PM protein content gradually decreased. Among all identified proteins, 12 showed significant differences in terms of their relative abundance. Increasing ubiquitin proteolysis and cytoskeleton proteins were observed from pre-véraison to post-véraison. Zeatin O-glucosyltransferase peaked at véraison, while ubiquitin-conjugating enzyme E2-21 was down-regulated at this stage. This proteome research provides the first information on PM protein characterization during the grape berry ripening process.
机译：葡萄（Vitis vinifera L. cv）的高纯度浆果质膜（PMs）。在浆果成熟的不同阶段，通过微粒体组分的两相分配分离出赤霞珠。通过二维电泳分离CHAPS和ASB-14的去污剂混合物可分离的PM蛋白。用银染检测到的2-D凝胶上的共生前浆果PM中的119个蛋白斑点进行了MALDI-TOF质谱分析。已鉴定出62个斑点为推定的PM蛋白，预测有1-6个跨膜螺旋，包括植物中报道的真实PM蛋白，例如ATP合酶，ABC转运蛋白和GTP结合蛋白。然后将它们分为八个功能类别，主要涉及运输，代谢，信号转导和蛋白质合成。鉴定出另外11个斑点为功能未知的蛋白质。检验和检验后样品分别在凝胶上染色98和86个斑点。在浆果成熟过程中，总PM蛋白含量逐渐降低。在所有鉴定出的蛋白质中，有12种在其相对丰度方面显示出明显差异。从检验前到检验后，观察到泛素蛋白水解和细胞骨架蛋白的增加。玉米蛋白O-葡萄糖基转移酶在突变期达到峰值，而泛素结合酶E2-21在此阶段下调。蛋白质组学研究提供了葡萄浆果成熟过程中PM蛋白表征的第一个信息。

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