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Journal articles
Identification of Leishmania-specific protein phosphorylation sites by LC-ESI-MS/MS and comparative genomics analyses
Abstract : Human pathogenic protozoa of the genus Leishmania undergo various developmental transitions during the infectious cycle that are triggered by changes in the host environment. How these parasites sense, transduce, and respond to these signals is only poorly understood. Here we used phosphoproteomic approaches to monitor signaling events in L. donovani axenic amastigotes, which may be important for intracellular parasite survival. LC-ESI-MS/MS analysis of IMAC-enriched phosphoprotein extracts identified 445 putative phosphoproteins in two independent biological experiments. Functional enrichment analysis allowed us to gain insight into parasite pathways that are regulated by protein phosphorylation and revealed significant enrichment in our data set of proteins whose biological functions are associated with protein turn-over, stress response, and signal transduction. LC-ESI-MS/MS analysis of TiO(2)-enriched phosphopeptides confirmed these results and identified 157 unique phosphopeptides covering 181 unique phosphorylation sites in 126 distinct proteins. Investigation of phosphorylation site conservation across related trypanosomatids and higher eukaryotes by multiple sequence alignment and cluster analysis revealed L. donovani-specific phosphoresidues in highly conserved proteins that share significant sequence homology to orthologs of the human host. These unique phosphorylation sites reveal important differences between host and parasite biology and post-translational protein regulation, which may be exploited for the design of novel anti-parasitic interventions.
Keywords :
Molecular Sequence Data
Leishmania
Life Cycle Stages
Mice
Amino Acid Sequence
Animals
Cells
Cultured
Chromatography
Liquid
Cluster Analysis
Databases
Protein
Electrophoresis
Gel
Two-Dimensional
Fungal Proteins
Humans
Phosphoproteins
Proteomics
Sequence Alignment
Spectrometry
Mass
Electrospray Ionization
Tandem Mass Spectrometry
https://hal-pasteur.archives-ouvertes.fr/pasteur-01433567
Contributor : Marie Martin Connect in order to contact the contributor
Submitted on : Thursday, January 12, 2017 - 5:10:01 PM
Last modification on : Thursday, January 6, 2022 - 2:50:02 PM
Contributor : Marie Martin Connect in order to contact the contributor
Submitted on : Thursday, January 12, 2017 - 5:10:01 PM
Last modification on : Thursday, January 6, 2022 - 2:50:02 PM
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