Evidence for heterotrimeric GTP-binding proteins in Toxoplasma gondii.
Toxoplasma gondii, an intracellular protozoan parasite, resides within a host-derived vacuole that is rapidly modified by a parasite-secreted membranous tubular network. In this study we investigated the involvement of heterotrimeric G proteins in the secretory pathway of T. gondii. Aluminum fluoride (AlFn), a specific activator of heterotrimeric G proteins, induced secretion from isolated tachyzoites of T. gondii in vitro, as seen by light optics and electron microscopy. In Western blot analyses, antibodies to G protein alpha subunits reacted with 39-42 kDa proteins from T. gondii isolates. Antibodies to G(o) alpha and Gs alpha coupled to the fluorescent probe fluorescein isothiocyanate localized to the paranuclear region of T. gondii. Gi3 alpha immunoprobes were confined to the cytoplasmic matrix of T. gondii and also labeled the parasitophorous vesicle. Fluorescein isothiocyanate-conjugated GA/1, an antipeptide antisera directed toward the GTP binding site common to G protein alpha subunits, was confined to the lateral cytoplasmic domain of the parasites where secretion is most prominent. In time-sequence studies using the GA/1 probe, the immunoreactive material shifted position during invasion of target cell to areas of active secretion.
Halonen
S K
SK
Department of Zoology and Physiology, Louisiana State University, Baton Rouge 70803, USA.
Weidner
E
E
Siebenaller
J F
JF
D017607:Aluminum Compounds / Q000494:pharmacology; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D002522:Chlorocebus aethiops; D019084:Fluorescent Antibody Technique, Indirect; D005459:Fluorides / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000032:analysis*; D015151:Immunoblotting; D015800:Protozoan Proteins / Q000032:analysis*; D051381:Rats; D014122:Toxoplasma / Q000737:chemistry* / Q000187:drug effects / Q000648:ultrastructure; D014617:Vacuoles; D014709:Vero Cells
D017607:Aluminum Compounds; D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D019204:GTP-Binding Proteins; D005459:Fluorides; C032311:aluminum fluoride