Bauw Guy, Nielsen Heidi V, Emmersen Jeppe, Nielsen Kåre L, Jørgensen Malene, Welinder Karen G
Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Denmark.
FEBS J. 2006 Aug;273(15):3569-84. doi: 10.1111/j.1742-4658.2006.05364.x.
The major potato tuber proteins of the Kuras cultivar, which is the dominant cultivar used in Northern Europe for industrial starch production, were analysed using 1D and 2D gel electrophoresis. The electrophoretic patterns varied significantly depending on the method of preparation and the potato variant (Solanum tuberosum). Proteins were characterized using MS and scored against potato protein databases, derived from both 'Kuras only' and 'all potato' expressed sequence tags (EST) and full-length cDNAs. Despite the existence of approximately 180 000 ESTs, the currently available potato sequence data showed a severe under-representation of genes or long transcripts encoding proteins > 50 kDa (3.5% of all) compared with the complete proteome of Arabidopsis thaliana (33% of all). We found that patatin and Kunitz protease inhibitor (KPI) variants are extraordinarily dominant in Kuras tuber and, most significantly, that their amino acid sequences are specific to Kuras. Other proteins identified include annexin, glyoxalase I, enolase and two lipoxygenases, the sequences of which are highly conserved among potato variants. Known S. tuberosum patatins cluster into three clades all represented in Kuras. S. tuberosum KPIs cluster into more diverse clades of which five were found in Kuras tuber, including a novel clade, KPI K, found to date only in Kuras. Furthermore, protein abundance was contrasted with the levels of corresponding gene transcripts found in our previous EST and LongSAGE studies of Kuras tuber.
库拉斯品种是北欧用于工业淀粉生产的主要马铃薯品种,使用一维和二维凝胶电泳对其主要块茎蛋白进行了分析。电泳图谱因制备方法和马铃薯变种(马铃薯)的不同而有显著差异。使用质谱对蛋白质进行表征,并与来自“仅库拉斯”和“所有马铃薯”表达序列标签(EST)及全长cDNA的马铃薯蛋白质数据库进行比对。尽管存在约180000个EST,但与拟南芥的完整蛋白质组(占全部的33%)相比,目前可用的马铃薯序列数据显示,编码大于50 kDa蛋白质的基因或长转录本严重不足(占全部的3.5%)。我们发现,马铃薯块茎蛋白和库尼茨蛋白酶抑制剂(KPI)变体在库拉斯块茎中极为占主导地位,最重要的是,它们的氨基酸序列是库拉斯特有的。鉴定出的其他蛋白质包括膜联蛋白、乙二醛酶I、烯醇化酶和两种脂氧合酶,它们的序列在马铃薯变种中高度保守。已知的马铃薯块茎蛋白聚为三个进化枝,在库拉斯中均有代表。马铃薯KPI聚为更多不同的进化枝,在库拉斯块茎中发现了其中五个,包括一个新的进化枝KPI K,迄今为止仅在库拉斯中发现。此外,还将蛋白质丰度与我们之前对库拉斯块茎进行的EST和LongSAGE研究中发现的相应基因转录本水平进行了对比。