Native DIGE for Quantitative and Functional Analysis of Protein Interactomes

Diksha Dani; Norbert A Dencher

doi:10.1007/978-1-0716-2831-7_4

Native DIGE for Quantitative and Functional Analysis of Protein Interactomes

Methods Mol Biol. 2023:2596:53-69. doi: 10.1007/978-1-0716-2831-7_4.

Authors

Diksha Dani^{1

2}, Norbert A Dencher³

Affiliations

¹ Institut für Biochemie und Biologie, Universität Potsdam, Potsdam-Golm, Germany.
² Physical Biochemistry, Department of Chemistry, Technische Universität Darmstadt, Darmstadt, Germany.
³ Physical Biochemistry, Department of Chemistry, Technische Universität Darmstadt, Darmstadt, Germany. norbert.dencher@physbiochem.tu-darmstadt.de.

PMID: 36378430
DOI: 10.1007/978-1-0716-2831-7_4

Abstract

Protein-protein interactions and multiprotein assemblies of water-soluble and membrane proteins are inherent features of the proteome, which also impart functional heterogeneity. One needs to consider this aspect while studying changes in abundance and activities of proteins in response to any physiological stimulus. Abundance changes in the components of a given proteome can be best visualized and efficiently quantified using electrophoresis-based approaches. Here, we describe the method of Blue Native Difference Gel Electrophoresis to quantify changes in abundance and activity of proteins in the context of protein-protein interactions. This method confers an additional advantage to monitor quantitative changes in membrane proteins, which otherwise is a difficult task.

Keywords: DIGE; Native PAGE; PolyacrylaKey wordsmide gel electrophoresis; Protein–protein interactions; Proteomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Electrophoresis, Gel, Two-Dimensional / methods
Membrane Proteins*
Proteome* / metabolism

Substances

Proteome
Membrane Proteins