Leishmania major: identification of developmentally regulated proteins in procyclic and metacyclic promastigotes |
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Authors: | Mojtahedi Zahra Clos Joachim Kamali-Sarvestani Eskandar |
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Affiliation: | a Department of Immunology, Shiraz University of Medical Sciences, Shiraz Medical School, P.O. Box 71345-1798, Shiraz 734-8553, Iran b Leishmaniasis Unit 1, Department of Medical Microbiology and Immunology, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany c Autoimmune Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran |
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Abstract: | The differentiation from procyclic to metacyclic promastigotes (metacyclogenesis) has been correlated with an increased infectivity in a number of Leishmania species. We compared the proteomes of procyclic and metacyclic promastigotes of L. major. Lysates from either life cycle stage were resolved by 2D-PAGE, followed by Coomassie brilliant blue staining. Spots were analyzed by MALDI-TOF MS. 25 protein spots were found to be differentially expressed during metacyclogenesis. We found that proteins involved in protein synthesis were less abundant in metacyclic promastigotes, while proteins involved in motility, including paraflagellar rod protein 1D, α-tubulin and β-tubulin were more abundant. Also, two mitochondrial enzymes (succinyl-CoA synthetase β subunit and cytochrome c oxidase subunit IV) were differentially expressed in both life cycle stages. Down-regulation of proteins related to synthetic pathway in metacyclic promastigotes is consistent with the arrested growth in this life cycle stage, while up-regulation of proteins related to motility in metacyclic promastigotes is in agreement with the high motility observed in this stage. |
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Keywords: | Trypanosomatid parasitic protozoa Promastigotes Procyclic Metacyclic Proteomics 2D-PAGE, two-dimensional polyacrylamide gel electrophoresis MALDI-TOF-MS, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry PNA&minus , peanut agglutinin IEF, isoelectric focusing MOWSE, molecular weight search (DTT), dithiothreitol |
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