Proteomic Analysis ofToxoplasma gondiiKI-1 Tachyzoites

Abstract

Abstract:We studied on the proteomic characteristics of Toxoplasma gondii KI-1 tachyzoites which were originally isolated from a Korean patient, and compared with those of the well-known virulent RH strain using 2-dimensional electrophoresis (2-DE), mass spectrometry, and quantitative real-time PCR. Two-dimensional separation of the total proteins isolated from KI-1 tachyzoites revealed up to 150 spots, of which 121 were consistent with those of RH tachyzoites. Of the remaining 29 spots, 14 showed greater than 5-fold difference in density between the KI-1 and RH tachyzoites at a pH of 5.0-8.0. Among the 14 spots, 5 from the KI-1 isolate and 7 from the RH strain were identified using MALDI-TOF mass spectrometry and database searches. The spots from the KI-1 tachyzoites were dense granule proteins (GRA 2, 3, 6, and 7), hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGRPTase), and uracil phosphoribosyltransferase (UPRTase). The spots from the RH strain were surface antigen 1 (SAG 1), L-lactate dehydrogenase (LDH), actin, chorismate synthase, peroximal catalase, hexokinase, bifunctional dihydrofolate reductase-thymidylate synthase (DHTR-TS), and nucleosidetriphosphatases (NTPases). Quantitative real-time PCR supported our mass spectrometric results by showing the elevated expression of the genes encoding GRA 2, 3, and 6 and UPRTase in the KI-1 tachyzoites and those encoding GRA 7, SAG 1, NTPase, and chorismate synthase in the RH tachyzoites. These observations demonstrate that the protein compositions of KI-1 and RH tachyzoites are similar but differential protein expression is involved in virulence.

Modulation of mouse macrophage proteome induced by Toxoplasma gondii tachyzoites in vivo

“…Rapid developments in proteomic techniques have provided novel approaches for proteomic studies of human and animal pathogens for elucidating the cellular responses toward infection, which will facilitate the identification of potential anti-infection drug targets and development of new diagnostic methods (Ye et al 2010). Several proteomic studies have been performed on T. gondii, which have helped to elucidate the specialized invasion organelles and their composition, as well as proteins associated with the cytoskeleton (Bradley et al 2005;El Hajj et al 2006;Kawase et al 2007;Fauquenoy et al 2008;Xia et al 2008;Ma et al 2009;Weiss et al 2009;Choi et al 2010;Che et al 2011). Nelson et al (2008 reported proteomic analysis of Toxoplasma-infected human foreskin fibroblasts (HFFs) in vitro, which indicated that the proteome of host cell responded in a dramatic way to T. gondii invasion.…”

Exploring Early and Late Toxoplasma gondii Strain RH Infection by Two-Dimensional Immunoblots of Chicken Immunoglobulin G and M Profiles

“…More than 163 and 150 protein spots from T . gondii strains RH and KI-1 tachyzoites, in corresponding order, were detected by 2-DE within a pH range of 3–10 after silver staining [ 32 ]. A high-resolution 2-DE gel separation over pH ranges 4–7 and 6–11 was also exploited to distinguish over 1000 polypeptides spots [ 20 ], and in a separate study, 1,227 protein spots of T .…”

The merits of designed ELISA avidity kit in detection of Toxoplasma gondii IgG antibody in laboratory conditions

“…[131415] Thus, in principle, ESA could play a role in the persistent stimulation of cell–mediated immunity in chronically infected healthy subjects. Excretory/secretory antigens may be the best form of antigens for stimulation of the cell-mediated immune response and a good candidate as a vaccine for toxoplasmosis prevention.…”