1 Introduction [1] [1] Drosophila melanogaster [2] Homo sapiens Mus musculus Gallus gallus Caenorhabditis elegans [3,4] [3] [2,5] C. elegans [6] In situ [3,4] Fig. 1 [6,7] Leishmania Trypanosoma N N Leishmania major Trypanosoma brucei Trypanosoma cruzi in vivo in vivo Leishmania 2 Materials and methods 2.1 PCR amplification and sub cloning LmPPEF Pfu For Rev LmPPEF LmPPEF For Rev For Rev Nde Bam TbPPEF T. brucei For Rev Nco Xho 2.2 Parasite culture, membrane fractionation and nucleic acid analysis L. major [8] 7 2 g g [8] 2.3 Antibody production and immunoblotting d Escherichia coli 2+ [9] [10] [8] 2.4 L. major LmPPEF For Rev Bam Eco [11] For For For Rev LmPPEF For Rev Nde Xba N Fig. 1 L. major [11] 2.5 Metabolic labelling and immunoprecipitation L. major [11] 3 3 2.6 Calcium mobility shift assay [12] 2 2.7 N [8,13] E. coli [8] [14] 3 2.8 Fluorescence microscopy L. major [15] l 6 3 Results 3.1 N N N N [8] L. major T. brucei [8,16,17] T. brucei in vitro T. brucei [18] N Leishmania T. brucei [11,14,19] N L. major T. brucei T. cruzi N [20] NMT Predictor N [21,22] Myristoylator [23] NMT Predictor [21] http://www.genedb.org/ N Myristoylator N Table 1 Tables S1–S3, supplementary data N L. major T. brucei T. cruzi T. cruzi [24] N [22,25] Table 1 Saccharomyces cerevisiae [21] N L. major [11] [14] T. brucei T. cruzi Tables S1–S3, supplementary data N T. brucei [26] L. major [19] Table 2 N 3.2 Kinetoplastid PPEF-like protein phosphatases [24,27] L. chagasi [28] T. cruzi [29] T. brucei [30] N LmjF12.0660 L. major Tb927.1.4050 T. brucei Tc00.1047053506529.380 Tc00.1047053510889.80 T. cruzi I L. major I T. brucei I T. cruzi [31] [32] Fig. 2 Fig. S1, supplementary data Fig. 2 Fig. 1 Fig. S2, supplementary data [33] Fig. S1, supplementary data D. melanogaster [1] Fig. S2, supplementary data T. cruzi [29] Plasmodium falciparum [34] Fig. S1, supplementary data [35] 2+ [1] N 3.3 L. major T. brucei [36] L. major T. brucei L. major T. brucei http://www.genedb.org/ [37] in silico L. major T. brucei Fig. 3 Fig. 3 L. major T. brucei Fig. 3 Fig. 1 L. major LmjF12.0660 Fig. 4 L. major Fig. 4 T. brucei 3.4 LmPPEF is not a calcium-binding protein 2+ Fig. 5 2+ L. major 2+ Fig. 5 L. major 2 Fig. 5 45 3.5 The N-termini of recombinant and wild type kinetoplastid PPEFs are substrates for acylation in vivo N Tables S1–S3, supplementary data E. coli [8] 3 Fig. 6 L. major T. brucei [8,14] in vivo L. major Fig. 1 N Fig. 1 Fig. 6 3 3 3 N Fig. 6 3 3 N [11] Fig. 6 in vivo N in vivo 3 Fig. 6 L. major Fig. 1 3 3.6 L. major L. major Fig. 7 Fig. 5 [38] Fig. 7 Fig. 7 L. major Fig. 6 Fig. 1 Fig. 7 N Fig. 7 Fig. 7 Fig. 7 L. major [11] N Leishmania [11,19 and this study] 4 Discussion in silico N L. major T. brucei in vivo [39,40] 2+ Drosophila [39,40] Fig. 5 Drosophila [41] [42] [43] TbPPEF T. brucei Appendix A Supplementary data doi:10.1016/j.molbiopara.2006.11.008 Appendix A Supplementary data