Proteome Profiling of Leishmania infantum Promastigotes
Proteome Profiling of Leishmania infantum Promastigotes
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DOI：
10.1111/j.1550-7408.2011.00549.x
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发表时间：
2011-07-01
影响因子：
2.2
通讯作者：
Larraga, Vicente
中科院分区：
文献类型：
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作者：
Alcolea, Pedro J.;Alonso, Ana;Larraga, Vicente
本文首次报道了婴儿利什曼原虫（MON-1 zymode）前鞭毛体阶段的蛋白质组学分析。在重复无菌培养物的早期对数期和稳定期制备总蛋白提取物，并通过2D电泳（pH 3-10）处理。基质辅助激光解吸/电离-串联飞行时间质谱共鉴定了28个差异调节蛋白。这种方法揭示了电子转移黄素蛋白（ETF）和真核细胞延伸因子1 α（eEF 1 α）亚基在蛋白质和mRNA水平上具有相同的差异表达模式，在稳定期上调。已检测到eEF 1 α亚基的低分子量同种型和替代加工形式。复制因子A的51 kDa亚基在对数前鞭毛体分裂中上调。这里描述的蛋白质都没有显示出与相应的mRNA水平相反的差异调节值。与以前的方法一起蛋白质组和转录组，这份报告有助于阐明的ETF，eEF 1 α亚基，40 S核糖体蛋白S12，α-微管蛋白和T-复合体蛋白1亚基γ在整个生命周期的寄生虫属利什曼原虫的差异调节模式。
A proteome analysis of the promastigote stage of the trypanosomatid parasite Leishmania infantum (MON-1 zymodeme) is described here for the first time. Total protein extracts were prepared at early logarithmic and stationary phases of replicate axenic cultures and processed by 2D electrophoresis (pH 3-10). A total of 28 differentially regulated proteins were identified by matrix-assisted laser desorption/ionization-tandem time of flight mass spectrometry. This approach has revealed that the electron transfer flavoprotein (ETF) and the eukaryotic elongation factor 1 alpha (eEF1 alpha) subunit have the same differential expression pattern at the protein and mRNA levels, up-regulation in the stationary phase. A low-molecular-weight isoform and an alternatively processed form of the eEF1 alpha subunit have been detected. A 51 kDa subunit of replication factor A is up-regulated in dividing logarithmic promastigotes. None of the proteins described here shows opposite differential regulation values with the corresponding mRNA levels. Taken together with previous approaches to the proteome and the transcriptome, this report contributes to the elucidation of the differential regulation patterns of the ETF, the eEF1 alpha subunit, the 40S ribosomal protein S12, alpha-tubulin and the T-complex protein 1 subunit gamma throughout the life cycle of the parasites from the genus Leishmania.