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Updated: Apr 26, 2026
Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
Published on: July 6, 2014
Identifying nuclear protein-protein interactions using GFP affinity purification and SILAC-based quantitative mass
H Irem Baymaz1, Cornelia G Spruijt, Michiel Vermeulen
1Department of Molecular Cancer Research, University Medical Center Utrecht, 85060, 3508 AB, Utrecht, The Netherlands.
Identifying protein-protein interactions (PPIs) is crucial for understanding protein function. This study presents a GFP affinity purification and SILAC mass spectrometry method to discover nuclear PPIs in mammalian cells.
Area of Science:
- Molecular Biology
- Proteomics
- Cell Biology
Background:
- Cellular proteins form complex structures, necessitating the study of protein-protein interactions (PPIs) for functional insights.
- Quantitative mass spectrometry techniques have advanced the unbiased identification of PPIs.
Purpose of the Study:
- To describe a method for identifying nuclear protein-protein interactions (PPIs) in mammalian cells.
- To detail a single-step GFP affinity purification combined with SILAC mass spectrometry for PPI discovery.
Main Methods:
- Utilizing a single-step Green Fluorescent Protein (GFP) affinity purification strategy.
- Employing Stable Isotope Labeling by Amino acids in Cell culture (SILAC)-based quantitative mass spectrometry.
- Applying the method to identify protein interactions within the nucleus of mammalian cells.
Main Results:
- The described method enables the identification of nuclear PPIs.
- This approach offers an efficient way to discover protein complexes.
Conclusions:
- The combined GFP affinity purification and SILAC mass spectrometry technique is effective for identifying nuclear PPIs.
- This method contributes to the understanding of protein complex formation and function in mammalian cells.
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