Furthermore, in motifs II and III, TbrPPX1 contains the sequence motifs DHN and DHH, respectively, which set it apart from the prune subfamily that contains the motifs DHH and DHR at the respective positions [8]. Characteristically, TbrPPX1 also
lacks the C-terminal extension of about 80 amino acids that is AP24534 purchase present in all vertebrate prunes, but is absent from the invertebrate prune homologues [9] and from the exopolyphosphatases. Figure CP673451 in vivo 1 TbrPPX1 is a predicted exopolyphosphatase that belongs to the subfamily 2 of the DHH superfamily. Dark boxes: motifs I – IV and V – VI of the DHH and the DHHA2 domains, respectively. Amino acid numbering corresponds to the TbrPPX1 sequence. Bold, underlined: active site motifs that discriminate the prune subfamily (DHH and DHR in motifs II and III, respectively) from the exopolyphosphatases/pyrophosphatases (DHN and DHH in motifs II and III, respectively). For a discussion of the functional consequences of his shift of the DHH signature from SGC-CBP30 motif II to motif III see [8]. Blast searching of the genomic databases of T. congolense, T. vivax, T. cruzi, L. major,
L. infantum, L. brasiliensis and L. tarentolae with TbrPPX1 demonstrated the presence of one orthologue of TbrPPX1 (three for T. cruzi) in each genome (Figure 2 and Table 1). The identical set of genes was also retrieved when searching the databases with the S. cerevisiae exopolyphosphatase ScPPX1 [GenBank: AAB68368]. All these TbrPPX1 homologues (group 1) share extensive sequence conservation and consist of about 380 amino acids, with calculated isoelectric points of about 5.5. For several of them, an exopolyphosphatase activity has been experimentally demonstrated [[14, 15], this study]. Figure 2 Neighbour distance tree of amino acid sequences of the kinetoplastid exo-and endopolyphosphatases. Group 1: cytosolic exopolyphosphatases; group: acidocalcisomal inorganic pyrophosphatases; group 3: pyrophosphatases. LY294002 For the designations of the individual genes and proteins see Table 1. Table 1 The exopolyphosphatases/pyrophosphatases of the kinetoplastids Organism GeneDB TrEMBL Gene ID Amino acids
Calc. MW Calc. pI Ref. Group 1 (exopoly-phosphatases) T. brucei Tb09.160.1950 (TbrPPX1) Q7Z032 3660027 383 42865 5.39 [16], this study T. congolense congo940f01.q1k_0 — 383 43004 5.66 T. cruzi Tc00.1047053504797.10 Q4DJ30 3545900 383 43029 5.95 15 Tc00.1047053511577.110 Q6Y656 383 43121 5.96 T. vivax tviv676c08.p1k_16 — 382 43434 5.68 L. braziliensis LbrM01_V2.0340 A4H355 5412361 387 42862 5.80 L. infantum LinJ01_V3.0310 A4HRF2 5066108 387 42626 5.59 L. major LmjF01.0310 Q25348 800604 388 42595 5.63 [14] L. tarentolae r1596.contig3320-2-1007-2215 — 387 43035 5.74 Group 2 (acidocalcisomal pyrophosphatases) T. brucei Tb11.02.4910 Q384W5 3665799 414 47330 5.73 [12, 13] Tb11.02.4930 Q7Z029 414 47307 5.70 T. cruzi Tc00.1047053511165.