Mapping protein complexes using covalently linked antibodies and isobaric mass tags
Antje Dittmann1, Sonja Ghidelli-Disse, Carsten Hopf
1Cellzome GmbH, Meyerhofstrasse 1, 69117, Heidelberg, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|May 6, 2014
Summary
This study introduces a method using immobilized antibodies for protein-protein interaction identification. It helps map protein complexes and networks under physiological conditions.
Area of Science:
- Proteomics
- Biochemistry
- Molecular Biology
Background:
- Quantitative proteomics and affinity enrichment are key for identifying protein-protein interactions.
- Understanding protein complexes and interaction networks is crucial in molecular biology.
Purpose of the Study:
- To describe an immunoaffinity enrichment method for identifying protein-protein interactions.
- To enable the study of endogenous proteins under near-physiological conditions.
Main Methods:
- Utilizing covalently immobilized antibodies for immunoaffinity enrichment.
- Employing shotgun mass spectrometry for protein identification.
- Using isobaric mass tag-based relative quantification for quantitative analysis.
Main Results:
- Successful identification of protein-protein interactions for endogenous proteins.
- Demonstration of the method's applicability under near-physiological conditions.
Conclusions:
- The described immunoaffinity enrichment approach is effective for unbiased protein-protein interaction identification.
- This technique facilitates the delineation of protein complexes and interaction networks.
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