Trypanosoma cruzi contains major pyrophosphate stores, and its growth in vitro and in vivo is blocked by pyrophosphate analogs.
High field (31)P nuclear magnetic resonance spectroscopy showed that inorganic pyrophosphate (P(2)O(7)(4-)) is more abundant than ATP in Trypanosoma cruzi, the causative agents of Chagas' disease. These results were confirmed by specific analytical assays, which showed that in epimastigotes, the concentrations of inorganic pyrophosphate and ATP were 194.7 +/- 25.9 and 37.6 +/- 5.5 nmol/mg of protein, respectively, and for the amastigote form, the corresponding concentrations were 358.0 +/- 17.0 and 36.0 +/- 1.9 nmol/mg of protein. High performance liquid chromatographic analysis of perchloric acid extracts of epimastigotes labeled for 3 h with (32)P-orthophosphate showed a significant incorporation of the precursor into inorganic pyrophosphate. Inorganic pyrophosphate was not uniformly distributed in T. cruzi but was shown by (31)P-NMR and chemical analysis to be particularly associated with acidocalcisomes, organelles shown previously to contain large amounts of phosphorus and various elements. Electron microscopy analysis of pyrophosphatase-treated permeabilized epimastigotes showed disappearance of the electron density of the acidocalcisomes. Nonmetabolizable analogs of pyrophosphate, currently used for the treatment of bone resorption disorders, selectively inhibited the proliferation of intracellular T. cruzi amastigotes and produced a profound suppression in the number of circulating trypomastigotes in mice with an acute infection of T. cruzi, offering a potentially new route to chemotherapy.
Urbina
J A
JA
Department of Chemistry, Department of Pathobiology, University of Illinois, Urbana-Champaign, Illinois 61802, USA.
Moreno
B
B
Vierkotter
S
S
Oldfield
E
E
Payares
G
G
Sanoja
C
C
Bailey
B N
BN
Yan
W
W
Scott
D A
DA
Moreno
S N
SN
Docampo
R
R
D000818:Animals; D011756:Diphosphates / Q000378:metabolism* / Q000494:pharmacology; D004164:Diphosphonates / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D008854:Microscopy, Electron; D000077268:Pamidronate; D013347:Subcellular Fractions / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*
D011756:Diphosphates; D004164:Diphosphonates; D000077268:Pamidronate