[Interaction between Toxoplasma GRA7 dense granule protein and the protein from host macrophages].
OBJECTIVE
To identify the protein from host macrophages which interacted with GRA7 dense granule protein of Toxoplasma gondii, and reveal the relationship between protein interaction and infection process.


METHODS
The recombinant GRA7 protein with N-terminal GST tag were used as a bait in in vitro GST Pull-down experiment, the proteins of THP-1 monocytic macrophage cell line were captured and identified by LC-MS/MS proteomics method. The in vivo protein interaction was verified by Co-IP experiment The overexpression of the target host protein by pcDNA3.1 (+) vector in THP-1 macrophage was further used to analyze the relationship between protein interaction and infection process.


RESULTS
The captured THP-1 cell protein was about Mr 29000, which was identified as human carbonic anhydrase 1 (hCA1). The significant in vivo protein-protein interaction between GRA7 and hCA1 was verified by Co-IP assay. The overexpression of hCA1 gene in THP-1 macrophage induced a higher propagation speed of Tgondii and the formation of the parasitophorous vacuole, but did nmt influence the number of the parasite.


CONCLUSIONS
There is a significant protein interaction between Toxoplasma GRA7 dense granule protein and hCA1 enzyme from host macrophages, which is positively related with the propagation speed of T. gondii.
Feng
F
Xue
Cai-ling
CL
Hong
Min-jun
MJ
Huang
Lan
L
Sun
Shao-bo
SB
Gan
Jun-chao
JC
Gu
D000953:Antigens, Protozoan / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D011994:Recombinant Proteins; D053719:Tandem Mass Spectrometry; D014122:Toxoplasma / Q000378:metabolism*
D000953:Antigens, Protozoan; C112573:GRA7 protein, Toxoplasma gondii; D015800:Protozoan Proteins; D011994:Recombinant Proteins