Identification and role of thiols in Toxoplasma gondii egress.
The nucleoside triphosphate hydrolase of Toxoplasma gondii is a potent apyrase that is secreted into the parasitophorous vacuole where it appears to be essentially inactive in an oxidized form. Recent evidence shows that nucleoside triphosphate hydrolase can be activated by dithiothreitol in vivo. On reduction of the enzyme, there is a rapid depletion of host cell ATP. Previous results also demonstrate a dithiothreitol induced egress of parasites from the host cell with a concurrent Ca2+ flux, postulated to be a consequence of the release of ATP-dependent Ca2+ stores within the tubulovesicular network of the parasitophorous vacuole. Reduction of the nucleoside triphosphate hydrolase appears crucial for its activation; however, the exact mechanism of reduction/activation has not been determined. Using a variety of techniques, we show here that glutathione promoters activate a Ca2+ flux and decrease ATP levels in infected human fibroblasts. We further show the in vitro activation of nucleoside triphosphate hydrolase by endogenous reducing agents, one of which we postulate might be secreted into the PV by T. gondii. Our findings suggest that the reduction of the parasite nucleoside triphosphate hydrolase, and ultimately parasite egress, is under the control of the parasites themselves.
Stommel
E W
EW
Department of Medicine, Dartmouth Medical School, Lebanon, New Hampshire 03756, USA. Elijah.W.Stommel@Hitchcock.org
Cho
E
E
Steide
J A
JA
Seguin
R
R
Barchowsky
A
A
Schwartzman
J D
JD
Kasper
L H
LH
D000111:Acetylcysteine / Q000494:pharmacology; D017766:Acid Anhydride Hydrolases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004396:Coloring Agents / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D043583:Nucleoside-Triphosphatase; D013438:Sulfhydryl Compounds / Q000502:physiology*; D014122:Toxoplasma / Q000187:drug effects / Q000378:metabolism*
D019086:Bridged Bicyclo Compounds, Heterocyclic; D004396:Coloring Agents; D013438:Sulfhydryl Compounds; C021149:bimanes; D000255:Adenosine Triphosphate; D017766:Acid Anhydride Hydrolases; D043583:Nucleoside-Triphosphatase; D002118:Calcium; D000111:Acetylcysteine