Institute of Plant Nutrition and Soil Science, Christian Albrechts University, Hermann-Rodewald-Strasse 2, 24118 Kiel, Germany.
Plant Methods. 2011 Dec 22;7:48. doi: 10.1186/1746-4811-7-48.
Proteins in the plant apoplast are essential for many physiological processes. We have analysed and compared six different infiltration solutions for proteins contained in the apoplast to recognize the most suitable method for leaves and to establish proteome maps for each extraction. The efficiency of protocols was evaluated by comparing the protein patterns resolved by 1-DE and 2-DE, and revealed distinct characteristics for each infiltration solution. Nano-LC-ESI-Q-TOF MS analysis of all fractions was applied to cover all proteins differentially extracted by infiltration solutions and led to the identification of 328 proteins in total in apoplast preparations. The predicted subcellular protein localisation distinguished the examined infiltration solutions in those with high or low amounts of intracellular protein contaminations, and with high or low quantities of secreted proteins. All tested infiltration solution extracted different subsets of proteins, and those implications on apoplast-specific studies are discussed.
质体蛋白在许多生理过程中必不可少。我们分析和比较了六种不同的质体蛋白提取液，以确定最适合叶片的提取方法，并为每种提取方法建立蛋白质组图谱。通过比较 1-DE 和 2-DE 解析的蛋白质图谱，评估了方案的效率，并发现每种提取液都有明显的特点。对所有馏分进行纳升 LC-ESI-Q-TOF MS 分析，以覆盖所有通过渗透液提取的差异蛋白质，总共鉴定到 328 种质体蛋白。预测的亚细胞蛋白定位将所检查的渗透液分为高或低细胞内蛋白污染、高或低分泌蛋白的渗透液。所有测试的渗透液提取的蛋白质都不同，讨论了这些对质体特异性研究的影响。
