TY - CHAP
T1 - Genome-Wide Proteomics and Phosphoproteomics Analysis of Leishmania spp. During Differentiation
AU - Pawar, Harsh
AU - Sathe, Gajanan
AU - Patole, Milind S.
N1 - Funding Information:
statement: Dr. Harsh Pawar is funded by an Israel Planning and Budgeting Commission (P.B.C.) fellowship.
Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2020.
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - Amastigote
KW - Leishmania development
KW - Parasite differentiation
KW - Promastigote
KW - Proteogenomics
KW - Signaling
KW - Tandem mass tag (TMT)
UR - http://www.scopus.com/inward/record.url?scp=85082519775&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-0294-2_12
DO - 10.1007/978-1-0716-0294-2_12
M3 - Chapter
C2 - 32221921
AN - SCOPUS:85082519775
SN - 9781071602935
T3 - Methods in Molecular Biology
SP - 161
EP - 176
BT - Trypanosomatids
A2 - Michels, Paul A. M.
A2 - Ginger, Michael L.
A2 - Zilberstein, Dan
PB - Humana Press Inc.
CY - New York
ER -