A proteomic analysis of arsenical drug resistance in Trypanosoma brucei.
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.
Foucher
Aude L
AL
Division of Infection and Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, UK.
McIntosh
Anne
A
Douce
Gill
G
Wastling
Jonathan
J
Tait
Andy
A
Turner
C Michael R
CM
D000818:Animals; D001152:Arsenicals / Q000494:pharmacology*; D004351:Drug Resistance; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D020543:Proteome / Q000032:analysis* / Q000187:drug effects / Q000235:genetics*; D014344:Trypanocidal Agents / Q000494:pharmacology; D014346:Trypanosoma brucei brucei / Q000187:drug effects* / Q000235:genetics
D001152:Arsenicals; D020543:Proteome; D014344:Trypanocidal Agents; C062789:melaminylthioarsenate