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 Scopus citations

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

ASJC Scopus subject areas

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

Fingerprint Dive into the research topics of 'Multiplexing strategy for simultaneous detection of redox-, phospho- and total proteome-understanding TOR regulating pathways in Chlamydomonas reinhardtii'. Together they form a unique fingerprint.

  • Cite this