Korean J Parasitol.  2006 Dec;44(4):303-312. 10.3347/kjp.2006.44.4.303.

Interactions between secreted GRA proteins and host cell proteins across the paratitophorous vacuolar membrane in the parasitism of Toxoplasma gondii

• Ahn HJ
• Kim S
• Kim HE
• Nam HW

• KMID: 1088762
• DOI: http://doi.org/10.3347/kjp.2006.44.4.303

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.

Keyword

Toxoplasma gondii; GRA proteins; parasitophorous vacuole; parasitophorous vacuole membrane; secretion; yeast two-hybrid technique; GRA-host protein interaction; parasitism

MeSH Terms

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