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Issue 17, 2015
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Multiplexing strategy for simultaneous detection of redox-, phospho- and total proteome – understanding TOR regulating pathways in Chlamydomonas reinhardtii

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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.

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

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Publication details

The article was received on 27 Feb 2015, accepted on 29 May 2015 and first published on 29 May 2015


Article type: Paper
DOI: 10.1039/C5AY00521C
Citation: Anal. Methods, 2015,7, 7336-7344
  • Open access: Creative Commons BY-NC license
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    Multiplexing strategy for simultaneous detection of redox-, phospho- and total proteome – understanding TOR regulating pathways in Chlamydomonas reinhardtii

    S. P. Rodrigues, S. Alvarez, E. G. Werth, W. O. Slade, B. Gau, E. B. Cahoon and L. M. Hicks, Anal. Methods, 2015, 7, 7336
    DOI: 10.1039/C5AY00521C

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