A quantitative proteomic and bioinformatics analysis of proteins in metacyclogenesis of Leishmania tropica

Abstract

Recently there has a growing interest in MS-based analysis on Leishmania for biology study, host-parasite interaction and drug target discovery. The aims of this study were to analyzed protein profiles in the procyclic and metacyclic stages of L. tropica, and investigate their potential role in metacyclogenesis molecular mechanisms. Sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH-MS) analysis was used to analyze protein profiles in each of procyclic and metacyclic stages. Proteins with a fold change>2 or <0.5 and p < 0.05 were considered to be significantly differentially expressed proteins (DEPs). The DEPs were subjected to gene ontology (GO), KEGG pathway and network analysis using PANTHER and STRING database, respectively. Quantitative real-time PCR of six selected genes validated the proteomic data. We quantified a total of 352 proteins in procyclic and metacyclic cells and 56 differentially expressed proteins (27 up/ 29down-regulated in metacyclic compared to procyclic). On the basis of biological processes in GO, the DEPs were primarily involved in ``metabolic process'' (GO: 0008152) and ``cellular process'' (GO: 0009987). In addition, several enriched GO terms were identified via molecular function, which among them ``catalytic activity'' (GO: 0003824) and ``binding'' (GO: 0005488) were disclosed as top category. The KEGG pathway analysis indicated ``metabolic pathways'' (p-value: 3.80E-08) including 17 genes term as the top pathway in DEPs. These findings bring a new insight in our understanding of the molecular characterization of metacyclogenesis and infective form in L. tropica. Comparative analysis of the proteome of both developmental stages of the L. tropica would help to the identification of proteins candidates for the development of new potential drug targets and vaccines.