Protein acetylation in pathogen virulence and host defense: In vitro detection of protein acetylation by radiolabeled acetyl coenzyme A

Authors: SCHREIBER, KARL - University Of California; Lewis, Jennifer

Submitted to: Methods in Molecular Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/24/2018
Publication Date: 5/1/2019
Citation: Schreiber, K.J., Lewis, J.D. 2019. Protein acetylation in pathogen virulence and host defense: In vitro detection of protein acetylation by radiolabeled acetyl coenzyme A. Methods in Molecular Biology. 1991:23-32. https://doi.org/10.1007/978-1-4939-9458-8_3.
DOI: https://doi.org/10.1007/978-1-4939-9458-8_3

Interpretive Summary: Protein networks are the machinery that carry out important functions of the cell. Proteins can be modified after synthesis (i.e. by acetylation), to help modulate their function. We developed a novel method to demonstrate that a protein has acetylation activity and to determine whether it can act on another protein. This protocol will allow laboratories studying many different areas of biology to apply this technique to their experimental systems. For instance, this method can be used to characterize proteins that are modified by bacterial pathogens during infection, or to ascribe a particular enzymatic activity to a protein of unknown function.

Technical Abstract: Protein acetylation has emerged as a common modification that modulates multiple aspects of protein function, including localization, stability, and protein-protein interactions. It is increasingly evident that protein acetylation significantly impacts the outcome of host-microbe interactions. In order to characterize novel putative acetyltransferase enzymes and their substrates, we describe a simple protocol for the detection of acetyltransferase activity in vitro. Purified proteins are incubated with 14C-acetyl CoA, separated electrophoretically, and acetylated proteins are detected by phosphorimaging or autoradiography.