A proteomic analysis of arsenical drug resistance in Trypanosoma brucei
A proteomic analysis of arsenical drug resistance in Trypanosoma brucei
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DOI：
10.1002/pmic.200500419
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发表时间：
2006-05-01
期刊：
影响因子：
3.4
通讯作者：
Turner, C. Michael R.
中科院分区：
文献类型：
--
作者：
Foucher, Aude L.;McIntosh, Anne;Turner, C. Michael R.
我们采用 2-DE 和 MS 来鉴定与布氏锥虫中砷耐药性相关的蛋白质。这种寄生虫会导致人类昏睡病，而砷耐药性是一个重大的潜在问题。对布氏锥虫药物敏感和耐药等基因系的可溶性蛋白质组中通过 2-DE 解析的约 2000 个斑点进行比较分析，确定了一个蛋白质斑点的缺失与砷药物 Cymelarsan 的耐药性相关。 MS将该蛋白与一对相同的串联基因Tb09.211.0120和0130相匹配，它们编码推定的新生多肽相关的复杂亚基。该蛋白质还以同种型的形式存在于耐药系和敏感系中，分子量相似，但 pi 不同。使用抗重组蛋白的抗体通过蛋白质印迹证实了同基因系之间的差异。药物敏感系和耐药系之间的两个基因序列相同，并且通过 RT-PCR 测定，两者均被转录。我们推测缺失的蛋白质亚型是由于缺乏 PTM 造成的。
We have undertaken 2-DE and MS to identify proteins associated with arsenical drug resistance in Trypanosoma brucei. This parasite causes sleeping sickness in humans, and arsenical drug resistance is a significant potential problem. Comparative analysis of approximately 2000 spots resolved by 2-DE in the soluble proteomes of drug-sensitive and drug-resistant isogenic lines of T. brucei identified a protein spot whose absence associated with resistance to the arsenical drug, Cymelarsan. MS matched this protein to an identical pair of tandem genes Tb09.211.0120 and 0130 that encode a putative nascent polypeptide associated complex subunit. This protein also occurs as an isoform located in both resistant and sensitive lines at a similar molecular weight, but different pi. The difference between isogenic lines was confirmed by Western blot using an antibody against recombinant protein. Both genes were identical in sequence between drug-sensitive and drug-resistant lines and both were transcribed as determined by RT-PCR. We postulate that the missing protein isoform arose due to the lack of a PTM.