Contents lists available at ScienceDirect Microbial Pathogenesis journal homepage: www.elsevier.com/locate/micpath Mitochondrial proteome profling of Leishmania tropica Minoo Tasbihi a , Faezeh Shekari b,d,∗∗ , Homa Hajjaran c , Leila Masoori a , Ramtin Hadighi a,∗ a Department of Medical Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran b Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran c Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran d Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran ARTICLE INFO Keywords: Leishmania Proteome Mitochondria LC-MS/MS ABSTRACT The mitochondrion of kinetoplastida has unique characteristics both in structure and function. To better un- derstand the mitochondrial proteome of the Leishmania tropica promastigote stage, liquid chromatography coupled with mass spectrometry (LC/MS/MS) approach was used. In the wake of mitochondria isolation and purity validation, 1212 proteins were identifed, among which approximately 44% of proteins belonged to the mitochondrial proteome. Several functions were enriched in mitochondrial proteome including tricarboxylic acid cycle and respiratory chain, protein folding, signalling, transport, lipid metabolism, amino acid, and nu- cleotide metabolism. Furthermore, the result of the present research was compared with the previous related studies. Gaining more information about vital metabolism of the cell and molecules can be used for therapeutic purposes. 1. Introduction Leishmania is a unicellular eukaryote, belonging to the protozoan order Kinetoplastida. The pathogenic Leishmania species cause diferent human diseases from cutaneous to visceral leishmaniasis in tropical and subtropical regions of the world [1]. L.tropica, the causative agent of anthroponotic cutaneous leishmaniasis (ACL) is endemic in various parts of Iran with a high incidence rate [2]. Drug-resistant L. tropica are now frequent in Iran [3]. Recently, a few cases of viscerotropic leishmaniasis caused by L. tropica has been reported in immunocompromised patients in Iran [4]. The genome of the Leishmania major (Friedlin strain) has been completely sequenced with 8272 protein-coding genes, 36% of which is recognized by a putative function [5]. These protozoa are among the primary eukaryotic species with mitochondria. The Leishmania mitochondria with other higher eu- karyotes are diferent both in number and volume. There are a large number of mitochondria in mammalian cells however, Leishmania has only one mitochondrion. The round single mitochondrion is composed of a compact matrix as well as the inner and outer membrane with many cristae. This organelle has a dense complex of DNA near the basal body called kinetoplast. Mitochondrial DNA contains maxicircles and minicircles, covering about 30% of the whole genome of the cell [6]. Most mitochondrial proteins are synthesized in the cytosol and have signal peptide for importing substances into mitochondria through specifc carriers in the inner and outer membranes [7]. The Leishmania mitochondria have shown characteristics which do not exist in other eukaryotes. Therefore, further studies about mitochondria are re- commended for therapeutic purposes on this unusual organelle [8]. The mitochondrion is the key site for energy production, along with other vital pathways such as cellular homeostasis and signalling, protein as well as lipid biosynthesis and beta-oxidation of fatty acids. It also plays a role in reactive oxygen species (ROS) production and apoptosis [8]. Proteomics analysis of mitochondria enriched by subcellular fractio- nation is a critical strategy for comprehensive identifcation of proteins in a specifc organelle. Previous studies have reported protein compo- sitions of mitochondria in yeast [9–11] as well as in mouse [12] and human tissues [13–15]. The comprehensive proteomic study of an en- riched Trypanosoma brucei mitochondria was performed by Panigrahi et al. [16], leading to the identifcation of 401 mitochondrial proteins with high confdence coupled with 615 mitochondrial and mitochon- dria-associated proteins. Here, we report the identifcation of the mi- tochondrial proteome of Leishmania tropica promastigotes by LC-MS/MS proteomics analysis. https://doi.org/10.1016/j.micpath.2019.103542 Received 5 February 2019; Received in revised form 17 April 2019; Accepted 14 May 2019 ∗ Corresponding author. ∗∗ Corresponding author. Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran E-mail addresses: faezehshekari@royaninstitute.org (F. Shekari), rhadighi@gmail.com (R. Hadighi). Microbial Pathogenesis 133 (2019) 103542 Available online 21 May 2019 0882-4010/ © 2019 Elsevier Ltd. All rights reserved. T