Proteomic Analysis of Toxoplasma gondii KI-1 Tachyzoites.
10.3347/kjp.2010.48.3.195
- Author:
Si Hwan CHOI
1
;
Tae Yun KIM
;
Sung Goo PARK
;
Guang Ho CHA
;
Dae Whan SHIN
;
Jong Yil CHAI
;
Young Ha LEE
Author Information
1. Department of Ophthalmology, Chungnam National University School of Medicine, Daejeon 301-747, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Toxoplasma gondii;
Korean Isolate-1 (KI-1);
proteomics;
quantitative real time PCR
- MeSH:
Electrophoresis, Gel, Two-Dimensional;
Gene Expression Regulation, Developmental;
Humans;
Molecular Sequence Data;
*Proteomics;
Protozoan Proteins/chemistry/*genetics/metabolism;
Toxoplasma/chemistry/*genetics/*growth & development/metabolism;
Toxoplasmosis/parasitology
- From:The Korean Journal of Parasitology
2010;48(3):195-201
- CountryRepublic of Korea
- Language:English
-
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.
