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Evaluation of proteome reference maps for cross-species identification of proteins by peptide mass fingerprinting

机译:蛋白质组参考图谱的评估,通过肽质量指纹图谱进行跨物种鉴定

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摘要

We tested whether proteome reference maps established for one species can be used for cross-specifies protein identification by comparing two-dimensional protein gel patterns and protein identification data of two closely related bacterial strains and four plant species. First, proteome profiles of two strains of the fully sequenced bacterium Sinorhizobium meliloti were compared as an example of close relatedness, high reproducibility and sequence availability. Secondly, the proteome profiles of three legumes (Medicago truncatula, Melilotus alba and Trifolium subterraneum), and the nonlegume rice (Oryza sativa) were analysed to test cross-species similarities. In general, we found stronger similarities in gel patterns of the arrayed proteins between the two bacterial strains and between the plant species than could be expected from the sequence similarities. However, protein identity could be concluded from their gel position, not even when comparing strains of the same species. Surprisingly, in the bacterial strains peptide mass fingerprinting was more reliable for species-specific protein identification than N-terminal sequencing. While peptide masses were found to be unreliable for cross-species protein identification, we present useful criteria to determine confident matching against species-specific expressed sequence tag databases. In conclusion, we present evidence that cautions the use of proteome reference maps and peptide mass fingerprinting for cross-species protein identification.
机译:我们通过比较二维密切相关的细菌菌株和四种植物物种的二维蛋白质凝胶模式和蛋白质识别数据,测试了为一种物种建立的蛋白质组参考图谱是否可用于交叉指定蛋白质识别。首先,比较了完全测序的两株苜蓿中华根瘤菌的两个菌株的蛋白质组谱,作为紧密相关性,高可重复性和序列可用性的一个例子。其次,分析了三种豆科植物(Medicago truncatula,Melilotus alba和Trifolium subterraneum)和非豆类水稻(Oryza sativa)的蛋白质组谱,以检验跨物种的相似性。通常,我们发现两个细菌菌株之间以及植物物种之间的阵列蛋白凝胶模式的相似性比序列相似性所预期的强。但是,即使在比较相同物种的菌株时,也不能从其凝胶位置推断出蛋白质同一性。出乎意料的是,在细菌菌株中,相比于N端测序,肽质量指纹图谱对物种特异性蛋白质鉴定更为可靠。虽然发现肽质量对于跨物种蛋白质鉴定不可靠,但我们提出了有用的标准来确定针对物种特异性表达序列标签数据库的可信匹配。总之,我们提出的证据提醒人们注意蛋白质组参考图谱和肽质量指纹图谱在跨物种蛋白质鉴定中的使用。

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