Multiplexing strategy for simultaneous detection of redox-, phospho- and total proteome-understanding TOR regulating pathways in Chlamydomonas reinhardtii

Silas P. Rodrigues, Sophie Alvarez, Emily G. Werth, William O. Slade, Brian Gau, Edgar B. Cahoon, Leslie M. Hicks

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

New methods for studying the complexity of multiple PTMs in functional proteomics are required to understand cell signaling processes. In this study, a multiplexing 2DE-based approach is introduced for parallel analysis of the redox-, phospho-, and total-proteome. This triplexing approach uses spectrally distinct fluorophores, is not matrix-specific and requires relatively low sample amounts with applicability to any cell/tissue type. This methodology was applied for the study of Target of Rapamycin (TOR) regulating pathways in Chlamydomonas reinhardtii. With emerging research demonstrating a complex yet unclear relationship between TOR kinase, autophagy, and lipid metabolism, rapamycin treatment was used to induce TOR inhibition in C. reinhardtii and redox-, phospho- and total proteome changes were assessed using the triplexing approach. We identified a total of 68 spot abundance changes in response to TOR inhibition which provide a basis for understanding this highly conserved, master regulator in algae.

Original languageEnglish (US)
Pages (from-to)7336-7344
Number of pages9
JournalAnalytical Methods
Volume7
Issue number17
DOIs
StatePublished - Sep 7 2015

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Sirolimus
Proteome
Multiplexing
Pulse time modulation
Cell signaling
Proteins
Fluorophores
Algae
Tissue
Phosphotransferases
Oxidation-Reduction
Proteomics
Lipid Metabolism

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemical Engineering(all)
  • Engineering(all)

Cite this

Multiplexing strategy for simultaneous detection of redox-, phospho- and total proteome-understanding TOR regulating pathways in Chlamydomonas reinhardtii. / Rodrigues, Silas P.; Alvarez, Sophie; Werth, Emily G.; Slade, William O.; Gau, Brian; Cahoon, Edgar B.; Hicks, Leslie M.

In: Analytical Methods, Vol. 7, No. 17, 07.09.2015, p. 7336-7344.

Research output: Contribution to journalArticle

Rodrigues, Silas P. ; Alvarez, Sophie ; Werth, Emily G. ; Slade, William O. ; Gau, Brian ; Cahoon, Edgar B. ; Hicks, Leslie M. / Multiplexing strategy for simultaneous detection of redox-, phospho- and total proteome-understanding TOR regulating pathways in Chlamydomonas reinhardtii. In: Analytical Methods. 2015 ; Vol. 7, No. 17. pp. 7336-7344.
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