Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes Using a Single-Step Affinity Purification

Marcin Luzarowski; Aleksandra Skirycz

doi:10.1007/978-1-0716-2624-5_8

Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes Using a Single-Step Affinity Purification

Methods Mol Biol. 2023:2554:107-122. doi: 10.1007/978-1-0716-2624-5_8.

Authors

Marcin Luzarowski¹, Aleksandra Skirycz²

Affiliations

¹ Zentrum für Molekulare Biologie der Universität Heidelberg, Heidelberg, Germany.
² Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany. as4258@cornell.edu.

PMID: 36178623
DOI: 10.1007/978-1-0716-2624-5_8

Abstract

Cellular protein-metabolite interactions (PMI), for decades relatively overlooked, are seeing a golden age in recent years. To facilitate simultaneous characterization of PMI and protein-protein interactions (PPI) of a given protein ("bait"), we developed a protocol that utilizes antibody-assisted affinity purification (AP) followed by liquid chromatography-mass spectrometry (LC-MS). Aside from its speed, simplicity, and adaptability to a variety of biological systems, its main strength lies in the parallel identification, in a near-physiological environment, of a given protein's protein and small-molecule partners.

Keywords: Affinity purification; Ligand; Mass spectrometry; Metabolites; Protein–metabolite interactions; Protein–protein interactions.

MeSH terms

Chromatography, Affinity / methods
Chromatography, Liquid
Mass Spectrometry / methods
Proteins* / chemistry

Substances

Proteins