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Identification and Quantitation of Signal Molecule-Dependent Protein Phosphorylation

Part of the Methods in Molecular Biology book series (MIMB,volume 1016)

Abstract

Phosphoproteomics is a fast-growing field that aims at characterizing phosphorylated proteins in a cell or a tissue at a given time. Phosphorylation of proteins is an important regulatory mechanism in many cellular processes. Gel-free phosphoproteome technique involving enrichment of phosphopeptide coupled with mass spectrometry has proven to be invaluable to detect and characterize phosphorylated proteins. In this chapter, a gel-free quantitative approach involving 15N metabolic labelling in combination with phosphopeptide enrichment by titanium dioxide (TiO2) and their identification by MS is described. This workflow can be used to gain insights into the role of signalling molecules such as cyclic nucleotides on regulatory networks through the identification and quantification of responsive phospho(proteins).

Key words

  • Gel-free proteomics
  • Mass spectrometry
  • Phosphoproteomics
  • Quantitation
  • TiO2 enrichment

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  • DOI: 10.1007/978-1-62703-441-8_9
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Groen, A., Thomas, L., Lilley, K., Marondedze, C. (2013). Identification and Quantitation of Signal Molecule-Dependent Protein Phosphorylation. In: Gehring, C. (eds) Cyclic Nucleotide Signaling in Plants. Methods in Molecular Biology, vol 1016. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-441-8_9

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  • DOI: https://doi.org/10.1007/978-1-62703-441-8_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-440-1

  • Online ISBN: 978-1-62703-441-8

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