Interactions between secreted GRA proteins and host cell proteins across the paratitophorous vacuolar membrane in the parasitism of Toxoplasma gondii.
10.3347/kjp.2006.44.4.303
- Author:
Hye Jin AHN
1
;
Sehra KIM
;
Hee Eun KIM
;
Ho Woo NAM
Author Information
1. Department of Parasitology and the Catholic Institute of Parasitic Diseases, College of Medicine, Catholic University of Korea, Seoul 137-701, Korea. howoo@catholic.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Toxoplasma gondii;
GRA proteins;
parasitophorous vacuole;
parasitophorous vacuole membrane;
secretion;
yeast two-hybrid technique;
GRA-host protein interaction;
parasitism
- MeSH:
Vacuoles/*metabolism;
Two-Hybrid System Techniques;
Toxoplasma/metabolism/*pathogenicity;
Protozoan Proteins/*metabolism/secretion;
Proteins/*metabolism;
Organelles/metabolism;
Intracellular Membranes/*metabolism;
Humans;
Hela Cells;
Gene Library;
Cytoplasmic Granules;
Animals
- From:The Korean Journal of Parasitology
2006;44(4):303-312
- CountryRepublic of Korea
- Language:English
-
Abstract:
Interactions between GRA proteins of dense granules in Toxoplasma gondii and host cell proteins were analyzed by yeast two-hybrid technique. The cMyc-GRA fusion proteins expressed from pGBKT7 plasmid in Y187 yeast were bound to host cell proteins from pGADT7-Rec-HeLa cDNA library transformed to AH109 yeast by mating method. By the selection procedures, a total of 939 colonies of the SD/-AHLT culture, 348 colonies of the X-alpha-gal positive and PCR, 157 colonies of the X-beta-gal assay were chosen for sequencing the cDNA and finally 90 colonies containing ORF were selected to analyze the interactions. GRA proteins interacted with a variety of host cell proteins such as enzymes, structural and functional proteins of organellar proteins of broad spectrum. Several specific bindings of each GRA protein to host proteins were discussed presumptively the role of GRA proteins after secreting into the parasitophorous vacuoles (PV) and the PV membrane in the parasitism of this parasite.
