Development of a safe, effective, and inexpensive
therapy for
African trypanosomiasis is an urgent priority. In this study, we evaluated the validity of Trypanosoma brucei
glycogen synthase kinase 3 (GSK-3) as a potential
drug target. Interference with the
RNA of either of two
GSK-3 homologues in bloodstream-form T. brucei parasites led to growth arrest and altered parasite morphology, demonstrating their requirement for cell survival. Since the growth arrest after RNA interference appeared to be more profound for T. brucei
GSK-3 "short" (Tb10.161.3140) than for T. brucei
GSK-3 "long" (Tb927.7.2420), we focused on T. brucei
GSK-3 short for further studies. T. brucei
GSK-3 short with an N-terminal
maltose-binding protein fusion was cloned, expressed, and purified in a functional form. The potency of a GSK-3-focused inhibitor library against the recombinant
enzyme of T. brucei
GSK-3 short, as well as bloodstream-form parasites, was evaluated with the aim of determining if compounds that inhibit
enzyme activity could also block the parasites' growth and proliferation. Among the compounds active against the cell, there was an excellent correlation between activity inhibiting the T. brucei
GSK-3 short
enzyme and the inhibition of T. brucei growth. Thus, there is reasonable genetic and chemical validation of
GSK-3 short as a
drug target for T. brucei. Finally, selective inhibition may be required for
therapy targeting the
GSK-3 enzyme, and a molecular model of the T. brucei
GSK-3 short
enzyme suggests that compounds that selectively inhibit T. brucei
GSK-3 short over the human
GSK-3 enzymes can be found.