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Updated: Jun 25, 2026

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
12:53

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics

Published on: July 6, 2014

SILAC for global phosphoproteomic analysis.

Genaro Pimienta1, Raghothama Chaerkady, Akhilesh Pandey

  • 1Department of Biological Chemistry, McKusick-Nathans Institute of Genetic Medicine, Baltimore, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 26, 2009
PubMed
Summary
This summary is machine-generated.

This study presents a quantitative phosphoproteomic strategy using stable isotope labeling with amino acids in cell culture (SILAC) and LC-MS/MS. This approach enables comprehensive identification and quantification of protein phosphorylation sites for cell signaling research.

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Area of Science:

  • Proteomics
  • Cellular Signaling
  • Biochemistry

Background:

  • Understanding protein phosphorylation patterns is crucial for cell signaling research.
  • Existing phosphoproteomic strategies often lack comprehensive quantitative information.
  • Identifying phosphorylation sites and their quantitative extent is a key goal.

Purpose of the Study:

  • To describe an experimental strategy for comprehensive phosphoproteomics.
  • To enable quantitative analysis of protein phosphorylation sites.
  • To highlight the importance of quantitative strategies in signal transduction.

Main Methods:

  • Stable Isotope Labeling with Amino acids in cell culture (SILAC) for quantitative labeling.
  • Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) for peptide and protein identification.
  • Integration of SILAC with LC-MS/MS for quantitative phosphoproteomics.

Main Results:

  • The described strategy allows for systematic and global elucidation of biological processes.
  • Quantitative data on phosphorylation site occupancy can be obtained.
  • Comprehensive identification of protein phosphorylation patterns is achievable.

Conclusions:

  • Quantitative phosphoproteomic strategies are essential for advancing cell signaling research.
  • The SILAC-coupled LC-MS/MS approach provides a robust platform for global phosphoproteome analysis.
  • This methodology facilitates a systematic understanding of biological signaling pathways.