br Materials and methods br Results br Discussion

Materials and methods
Results
Discussion Proteomics analyses generate mountains of data, expression information for hundreds or thousands of proteins in a single experiment (Wojcik and Schächter, 2000). Recently proteomics and bioinformatics provide new understandings about the processes in living cells and organisms (Darabi and Farhadi-Nejad, 2013, Darabi and Seddigh, 2015, Seddigh and Darabi, 2014, Seddigh and Darabi, 2015, Seddigh and Darabi, 2016). Bioinformatics has been used to study the conserved motifs and structures of proteins (Baloğlu et al., 2011, Bordoli and Schwede, 2012, Zhang et al., 2012). In this study, bioinformatics tools were used to identify the characteristics of two families of P-ATPases including P-ATPase 13A1 and 13A3 families, in different insect species. P-ATPases are a large family of integral membrane transporters that are of vital importance in all kingdoms of lives. The superfamily of P-ATPases includes membrane-bound enzymes which utilize the potential energy of ATP for CUDC-907 of ions (Hatori et al., 2008, Mandal et al., 2002, Pilankatta et al., 2009, Pilankatta et al., 2011). Members of this family generate and maintain essential (electro-) chemical gradients across cellular membranes, by translocating cations, heavy metals and lipids. Availability of structural information on P-type ATPases has increased considerably within recent years. This has led to the elucidation of hallmark intermediate conformations along the reaction cycle, so providing the structural basis for ATP hydrolysis and ion transport (Bublitz et al., 2010). Motif analysis of P-ATPase families was done in order to find patterns of conserved motifs by MEME program. Finding a cluster of numerous, similar subsequences in a set of biopolymer sequences is evidence that the subsequences occur not by chance because they share some biological function which accounts for similarity of a subset of subsequences might be a common protein binding site or splice junction in the case of DNA sequences, or active site of related enzymes in the case of protein sequences (Bailey and Elkan, 1994). Ten motifs were identified in each family of P-ATPase which their sequence logos are shown in Fig. 1. Motif analysis of P-ATPase 13A1 family showed all ten motifs were present in all insect species but motif 10, which was not observed in Halyomorpha halys. In P-ATPase 13A3 family, ten conserved motifs were common in 28 insect species. In this protein family, motif 9 with 50aa was not observed in Atta cephalotes while motifs 4 and 5 with 50aa were not identified in Trichogramma pretiosum (Table 3). Primary structural analysis showed that the most total number of atoms and the most molecular weight of P-ATPase13A1 family belonged to Musca domestica with 19,426 and 136,897.8Da, respectively. While in P-ATPase13A3 family these most values were observed in B. terrestris with 19,801 atoms and the molecular weight about 139,917.2Da. According to these analyses in P-ATPase13A1 and P-ATPase 13A3 family the least number of atoms and molecular weight were found in Halyomorpha halys and Acyrthosiphon pisum, respectively. Isoelectric Point is a pH at which a protein carries no net charge. Below the isoelectric point proteins carry a net positive charge and above it a net negative charge. The isoelectric point is of significance in protein purification because it is the pH at which solubility is often minimal and at which mobility in an electrofocusing system is zero, and therefore the point at which the protein will accumulate. In computing PI analysis the most isoelectric point in P-ATPase13A1 family belonged to Bt-13A1 (8.26) whereas the least was found in Hh-13A1 (6.61), while in the other family they were observed in Bt-13A3 and Ap-13A3, respectively. This analysis revealed that P-ATPase proteins are different from each other, so that they may be acidic or alkaline in nature.