Genome-Wide Proteomics and Phosphoproteomics Analysis of Leishmania spp. During Differentiation

Harsh Pawar (Lead / Corresponding author), Gajanan Sathe, Milind S. Patole

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Determining variations in protein abundance and/or posttranslational modification as a function of time or upon induction by a signal in a particular cell type is central to quantitative proteomics. Isobaric labeling methodologies now allow for parallel quantification of proteins at various conditions concurrently or multiplexing in relatively quantitative proteomics workflows. Hence, mapping the protein expression profiles of various developmental stages of Leishmania parasites is possible with high-resolution mass spectrometry. To analyze global changes in protein expression and cellular signaling pathways during Leishmania differentiation and development is possible with a quantitative proteomics approach. The tandem mass tags (TMT) approach provides a chemical labeling method based on the principle of amine reactive tags; the maximum number of conditions that can be multiplexed is 10-plex. We describe herein a detailed method for sample preparation, TMT-labeling, mass spectrometry and data analysis of different developmental stages of Leishmania donovani parasites. This quantitative proteomic approach is useful to study dynamic changes in protein expression levels during L. donovani differentiation, and also allows in-depth analysis of signaling pathways via phosphoproteomics.

Original languageEnglish
Title of host publicationTrypanosomatids
EditorsPaul A. M. Michels, Michael L. Ginger, Dan Zilberstein
Place of PublicationNew York
PublisherHumana Press Inc.
Pages161-176
Number of pages16
ISBN (Electronic)9781071602942
ISBN (Print)9781071602935
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2116
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Amastigote
  • Leishmania development
  • Parasite differentiation
  • Promastigote
  • Proteogenomics
  • Signaling
  • Tandem mass tag (TMT)

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