Evidence for heterotrimeric GTP-Binding proteins in Toxoplasma gondii

被引:10
作者
Halonen, SK [1 ]
Weidner, E [1 ]
Siebenaller, JF [1 ]
机构
[1] LOUISIANA STATE UNIV, DEPT ZOOL & PHYSIOL, BATON ROUGE, LA 70803 USA
关键词
aluminum fluoride activation; antibody labeling; exocytosis; immunocytochemical probes; parasitophorous vacuoles; tubule secretion;
D O I
10.1111/j.1550-7408.1996.tb01389.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 (AlF(a)lpha), 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 ct subunits reacted with 39-42 kDa proteins from T. gondii isolates. Antibodies to G(o alpha) and G(s alpha) coupled to the fluorescent probe fluorescein isothiocyanate localized to the paranuclear region of T. gondii. G(i3 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.
引用
收藏
页码:187 / 193
页数:7
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