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Proteomic Analysis of Toxoplasma gondii KI-1 Tachyzoites

Korean Journal of Parasitology 2010년 48권 3호 p.195 ~ 201
최시환 ( Choi Si-Hwan ) - 충남대학교 의과대학 안과학교실

 ( Kim Tae-Yun ) - 충남대학교 의과대학 감염생물학교실 의학연구소
 ( Park Sung-Goo ) - Korea Research Institute of Bioscience and Biotechnology Medical Proteomics Research Laboratory
차광호 ( Cha Guang-Ho ) - 충남대학교 의과대학 감염생물학교실 의학연구소
신대환 ( Shin Dae-Whan ) - 충남대학교 의과대학 감염생물학교실 의학연구소
채종일 ( Chai Jong-Yil ) - 서울대학교 의과대학 기생충학교실
이영하 ( Lee Young-Ha ) - 충남대학교 의과대학 감염생물학교실 의학연구소

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 nucleoside-triphosphatases (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.

키워드

Toxoplasma gondii; Korean Isolate-1 (KI-1); proteomics; quantitative real time PCR
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