edoc

Measurement of in vivo protein binding affinities in a signaling network with mass spectrometry

Gencoglu, Mumun and Schmidt, Alexander and Becskei, Attila. (2017) Measurement of in vivo protein binding affinities in a signaling network with mass spectrometry. ACS Synthetic Biology, 6 (7). pp. 1305-1314.

[img] PDF - Accepted Version
850Kb

Official URL: http://edoc.unibas.ch/54752/

Downloads: Statistics Overview

Abstract

Protein interaction networks play a key role in signal processing. Despite the progress in identifying the interactions, the quantification of their strengths lags behind. Here we present an approach to quantify the in vivo binding of proteins to their binding partners in signaling-transcriptional networks, by the pairwise genetic isolation of each interaction and by varying the concentration of the interacting components over time. The absolute quantification of the protein concentrations was performed with targeted mass spectrometry. The strengths of the interactions, as defined by the apparent dissociation constants ranged from subnanomolar to micromolar values in the yeast galactose signaling network. The weak homodimerization of the Gal4 activator amplifies the signal elicited by glucose. Furthermore, combining the binding constants in a feedback loop correctly predicted cellular memory, a characteristic network behavior. Thus, this genetic-proteomic binding assay can be used to faithfully quantify how strongly proteins interact with proteins, DNA and metabolites.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Computational & Systems Biology > Synthetic Microbiology (Becskei)
UniBasel Contributors:Becskei, Attila and Schmidt, Alexander
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:American Chemical Society
e-ISSN:2161-5063
Note:Publication type according to Uni Basel Research Database: Journal article
Language:English
Identification Number:
Last Modified:13 Apr 2018 12:22
Deposited On:02 Oct 2017 12:31

Repository Staff Only: item control page