Interestingly, a recent study showed that the select ive inhibition of GADD34, a human regulator of PP1, by guanabenz www.selleckchem.com/products/Bortezomib.html was able to restore proteostasis and to protect stressed cells. This further confirms that interfering with the interaction of PP1 regulators and or dissociation of the comple can help to better understand the role of PfPP1 and to create new means to develop antimalarials. Methods Genome databases searches and sequences analysis Putative Inhibitor 2 sequences were searched using BLASTp on sequences available in GenBank, PlasmoDB, To oDB, SchistoDB, enbase and OrthoMCLDB databases. The human I2 Plasmo dium falciparum I2 sequences alignment was performed using the ClustalW program and was manually corrected.

Phylogenetic analyses and secondary structure prediction Protein sequences were aligned using the ClustalW algorithm implemented in the BioEdit v7. 1 software, and manually corrected. Ma imum likehood trees were built using MEGA5 under the JTT I G model, with 100 boot strap repetitions. of the following species Plasmodium falciparum, Plasmodium berghei, Plasmodium chabaudi, Plasmodium knowlesi, Plasmodium viva , Plasmodium yoeli yoeli, To oplasma gondii, Arabidopsis thaliana, Homo sapiens, Mus musculus, Trypanosoma brucei, Tetrahymena thermophila, enopus laevis, Danio rerio, Saccharomyces cerevisiae, Theileria parva, Drosophila melanogaster, Leishmania major, Oryza sativa, cae norhabditis elegans and Schistosoma mansoni. PfI2 secondary prediction was carried out using the PsiPred software and the potential nuclear signal localization was performed using the PSORTII soft ware.

Plasmids Plasmids pCR2. 1 TOPO, pQE30, pGE 4T3, pETDuet 1, pGADT7 and pGBKT7 were purchased from Invitrogen, Qiagen, Life Sciences, Novagen and Clontech respect ively. Plasmid pCAM HA, and pCAM were kind gifts of Dr C. Doerig. Monoclo nal anti HA and anti Myc antibodies were purchased from Roche and Invitrogen respectively. Preparation of parasites P. falciparum 3D7 and HB3 clones were grown according to Trager and Jensen, in RPMI 1640 medium sup plemented with 0. 5% AlbuMA II, 0. 2 mM Hypo anthin and 20 ug ml Gentamycin, in the presence of O erythrocytes. Parasites were synchronized by a double sorbitol treatment as previously described. In order to isolate total RNA or proteins, parasitized erythrocytes were prepared by saponin AV-951 lysis and either resuspended in Trizol or in phosphate buffered saline containing EDTA free protease inhibitor cocktail. For some e periments, infected red blood cells were purified using Percoll sorbitol density gradients with slight modifications.