Journal
PROTEOMICS
Volume 6, Issue 9, Pages 2726-2732Publisher
WILEY
DOI: 10.1002/pmic.200500419
Keywords
cymelarsan; drug resistance; parasite; proteome; Trypanosoma brucei
Funding
- Medical Research Council [G9722968] Funding Source: Medline
- Wellcome Trust Funding Source: Medline
- Medical Research Council [G9722968] Funding Source: researchfish
- MRC [G9722968] Funding Source: UKRI
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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.
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